FOWL ADENOVIRUS 9 (FADV-9) VECTOR SYSTEM AND ASSOCIATED METHODS

Abstract

Described are recombinant viral vectors obtained from fowl adenovirus 9 (FAdV-9) and associated methods. The recombinant FAdV-9 vectors may include one or more deletions at the left end and/or right end of the FAdV-9 genome. Optionally, the vectors include one or more exogenous nucleotide sequences, such a sequences encoding for a polypeptide of interest. The recombinant FAdV-9 vectors may be used as a dual delivery vector. Also described is the use of the vectors for generating an immunogenic response in a subject and/or for the prevention of disease.

Description

RELATED APPLICATION

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62/190,913 filed Jul. 10, 2015, the contents of which are hereby incorporated by reference in their entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to the fowl adenovirus 9 (FAdV-9) and more particularly to a FAdV-9 dual delivery vector system and associated methods as well as use of the same for the prevention of disease.

BACKGROUND OF THE INVENTION

[0003] Fowl adenoviruses (FAdVs) are ubiquitous poultry pathogens and are members of the family Adenoviridae.

[0004] Adenoviruses (AdVs) of the genus Mastadenovirus have been examined as anti-cancer agents (Huebner et al. (1956), Cody & Douglas (2009), Yamamoto & Curiel (2010)) and vaccine vectors (Lasaro & Ertl (2009)).

[0005] The problem of preexisting immunity against HAdV-5, exemplified in the STEP HIV trial that employed recombinant HAdV-5 (Buchbinder et al. (2008), McElrath et al. (2008)), has generated interest in the development of less common AdV serotypes and nonhuman AdVs as both oncolytic ((Cody & Douglas (2009), Gallo et al. (2005), Shashkova et al. (2005)) and vaccine vectors (Barouch (2008), Lasaro & Ertl, (2009), Sharma et al. (2009)). Fowl adenoviruses (FAdVs) of the genus Aviadenovirus, including species FAdV-A to FAdV-E (Adair, B. & Fitzgerald, S. (2008), Benko et al (2005)), are being developed as vaccine vectors. The first generation of FAdV-based vaccine vectors have proven to be effective at eliciting an antibody response against a delivered transgene (Corredor & Nagy (2010b), Ojkic & Nagy (2003)), and in chickens have conferred protective immunity against infectious bursal disease virus (IBDV) (Francois et al. (2004), Sheppard et al. (1998)) and infectious bronchitis virus (Johnson et al. (2003)). Analysis of the complete genomes of FAdV-1, the chicken embryo lethal orphan (CELO) virus (Chiocca et al. (1996)), and FAdV-9 (Ojkic & Nagy (2000)) (species FAdV-A and FAdV-D, respectively), and the terminal genomic regions of FAdV-2, -4, -10, and -8 (Corredor et al. (2006), Corredor et al. (2008)) has shown that the FAdVs share a common genome organization.

[0006] Adenovirus-based vaccine vectors have proven to be promising tools for controlling pathogens (Bangari & Mittal (2006), Ferreira et al. (2005)). The first generation of fowl adenovirus (FAdV) based vaccine vectors have been effectively used to induce an antibody response against an inserted foreign gene (transgene) (Corredor, & Nagy (2010a), Ojkic & Nagy (2003)), and in chickens have conferred protective immunity against infectious bursal disease virus (Francois et al. (2004), Sheppard et al. (1998)) and infectious bronchitis virus (Johnson et al. (2003)).

[0007] Current state of the art adenoviral vectors, especially fowl adenoviruses, are hampered by the limit of the size in the foreign DNA insert size in the vector DNA. Consequently it is not possible to clone in particularly large DNA inserts representing important parts of immunogenic proteins into an independently replicating virus. Nor it is possible to clone in more than one gene. The value of having a vector capable of expressing dual or even multivalent antigens is that only one vaccine would be needed to protect against two or more diseases. This is not possible with the current state of the art adenoviral vectors due to lack of cargo space.

[0008] Therefore, there remains a need for novel adenoviral vectors and in particular for novel dual delivery adenoviral vectors and uses thereof.

SUMMARY OF THE INVENTION

[0009] The inventors have developed novel adenoviral vectors based on recombinant fowl adenovirus 9 (FAdV-9). The vectors are particularly useful for the delivery and/or expression of exogenous sequences and as dual delivery adenoviral vectors. Inserted into the novel vector can be one or more exogenous nucleotide sequences. Optionally, the one or more exogenous nucleotide sequences code for one or more antigenic sites of a disease of concern.

[0010] Accordingly, in one aspect, there is provided a recombinant fowl adenovirus 9 (FAdV-9) viral vector. In one embodiment, the recombinant FAdV-9 viral vector is a dual delivery viral vector.

[0011] In one aspect, the FAdV-9 viral vector has a deletion at the left end of the genome. In one embodiment, the deletion at the left end of genome comprises a deletion of one or more of ORF0, ORF1 and ORF2. In one embodiment, the FAdV-9 viral vector has a deletion at the right end of the genome. In one embodiment, the deletion at the right end of the genome comprises a deletion of one or more of ORF19, TR2, ORF17 and ORF11. In one embodiment, the FAdV-9 viral vector has deletions at both the left end and right end of the genome. In one embodiment, the FAdV-9 viral vector has a deletion of ORF0, ORF1, ORF2, TR2, ORF17 and ORF11. In one embodiment, the FAdV-9 viral vector has a deletion of ORF1, ORF2, TR2, ORF17 and ORF11. In other embodiment, the FAdV-9 viral vector has a deletion ORF1, ORF2, and ORF 19. In other further embodiment, the FAdV9-viral vector has a deletion of ORF1, ORF2, and ORF19, TR2 or ORF11.

[0012] In one embodiment, the FAdV-9 viral vector has a deletion at the left end of the genome of about 2291 base pairs, optionally between about 1900 and 2500 base pairs. In one embodiment, the FAdV-9 viral vector has a deletion at the right end of the genome of about 3591 base pairs. In one embodiment, the FAdV-9 viral vector has a deletion at the right end of the genome of between about 3000 base pairs and 4000 base pairs.

[0013] In one embodiment, the viral vector comprises a nucleotide sequence with at least 80%, at least 90%, at least 95%, at least 98% or at least 99% sequence identity to the sequence with the two deletions shown in FIG. 10A. In one embodiment, the viral vector comprises or consists of the nucleotide sequence shown in SEQ ID NO: 1 with one or more deletions, optionally one or more deletions shown in FIG. 10A.

[0014] In one embodiment, the viral vector has an insert capacity of greater than 4000 bp, greater than 5000 bp, greater than 6000 bp or optionally greater than 7000 bp.

[0015] In one embodiment, the viral vector comprises at least one of the following:

[0016] (a) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 575 to 2753 have been deleted,

[0017] (b) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 847 to 2753 have been deleted,

[0018] (c) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 42,398 have been deleted,

[0019] (d) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 34,220 to 36,443 have been deleted,

[0020] (e) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 40,561 have been deleted or wherein nucleotides 41,461 to 42,398 have been deleted, and

[0021] (f) a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity to the nucleotide sequences set out in (a), (b), (c), (d) or (e).

[0022] For example, in one embodiment, the viral vector comprises, consists essentially of, or consists of a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of nucleotides 575 to 2753, 847 to 2753, and/or 38,807 to 42,398 have been deleted. In one embodiment, the viral vector comprises, consists essentially of, or consists of a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part nucleotides 575 to 2753 and 38,807 to 42,398 have been deleted. In one embodiment, the viral vector comprises, consists essentially of, or consists of a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part nucleotides 847 to 2753 and 38,807 to 42,398 have been deleted. In another embodiment, the viral vector comprises, consists essentially of, or consists of a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part nucleotides 847 to 2753 and 34,220 to 36,443 have been deleted. In another embodiment, the viral vector comprises, consists essentially of, or consists of a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part nucleotides 847 to 2753, 34,220 to 36,443, and 38,807 to 40,561 or 41,461 to 42398 have been deleted.

[0023] Another aspect of the invention considers a viral vector having inserted one or more exogenous nucleotide sequences. In one embodiment, the viral vector comprises one or more exogenous nucleotide sequences coding for one or more polypeptides of interest, optionally one or more antigenic and/or therapeutic polypeptides. In one aspect, the viral vector is a dual delivery vector capable of expressing two or more exogenous nucleotide sequences. In one embodiment, the viral vector comprises exogenous nucleotide sequences coding for one or more antigenic sites of a disease of concern. In one embodiment, the viral vector comprises a sequence corresponding to at least one gene listed in Table 1 or a homolog thereof.

[0024] Also provided are host cells transformed with one or more viral vectors as described herein.

[0025] A further aspect of the invention is a method for producing a viral vector as described herein. In one embodiment, the method comprises inserting an exogenous nucleotide sequence into a recombinant FAdV-9 viral vector as described herein.

[0026] In one embodiment, the FAdV-9 viral vector comprises one or more recombinant control sequences, such as one or more promoters. Optionally, the viral vector comprises one or more cloning sites to facilitate recombinant insertion of exogenous nucleotide sequences into the viral vector.

[0027] In one aspect, there is provided an immunogenic composition comprising aFAdV-9 viral vector as described herein having an exogenous nucleotide sequence coding for at least one antigenic site of a disease of concern inserted therein. In one embodiment, the immunogenic composition further comprises a pharmaceutically acceptable carrier. In one embodiment, the immunogenic composition further comprises an adjuvant. In one embodiment, the immunogenic composition is a vaccine.

[0028] In another aspect, there is provided a method for generating an immunogenic response in a subject. In one embodiment, the method comprises administering to the subject a viral vector or immunogenic composition as described herein. Also provided is a viral vector or immunogenic composition as described herein for use in generating an immunogenic response in a subject. In one embodiment, the methods and uses described herein are for generating an immunogenic response against a disease antigen, optionally one or more diseases listed in Table 1. In one embodiment, the methods and uses described herein are for the prevention of disease and/or vaccinating a subject against one or more diseases.

[0029] Other features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples while indicating preferred embodiments of the invention are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from the detailed description.

BRIEF DESCRIPTION OF THE FIGURES

[0030] The novel features of the present invention are established particularly in the appended claims. However, the invention itself together with other objects and advantages thereof will be better understood in the following detailed description of a specific embodiment, when read along with the appended figures, in which:

[0031] FIG. 1 shows a FAdV-9 vector system (FAdmid). The Fadmid has the following characteristics: Non-pathogenic strain of FAdV-9, dispensable regions at left and right end, recombinant FAdVs grow like wild-type FAdV-9, decreased virus shedding and antibody response to viral backbone and EGFP (reporter gene) is expressed using this backbone when exogenous (foreign) promoter (CMV) is used (Corredor, J. C. & Nagy, E. (2010b)).

[0032] FIG. 2 shows the left end sequence of FAdV-9. ORF1 and 1C function: Modulates the innate and adaptive immune response; ORF0, 1A, 1B and 2 function: Unknown. When ORF0 is deleted the native early promoter is not functioning--no foreign gene expression (e.g. m-Cherry). When only ORF 1 and 2 are deleted the native promoter works.

[0033] FIG. 3 shows the deletion of ORFs 0, 1 and 2. (A) Flowchart of the generation of pFAdV9-.DELTA.0-1-2-RED: ORF0, ORF1, 1A, 1B, 1C and 2 were replaced with the CAT cassette flanked on both sides with SwaI sites to generate pFAdV9-.DELTA.0-1-2CAT by homologous recombination. The CAT cassette was removed by SwaI digestion followed by religation to generate unmarked pFAdV9-.DELTA.0-1-2SwaI. The mCherry coding sequence was then cloned into SwaI site to generate pFAdV9-.DELTA.0-1-2RED, MLP, major late promoter; ITR, inverted terminal repeat; (B) and (D) NotI digestions and PCR amplification of DNA to verify the FAdmids and the corresponding viruses. Lanes .DELTA.0-2; pFAdV9-.DELTA.0-1-2RED or passage 3 virus; lanes wt: pFAdV9-wt or virus; M: 1 kb DNA ladder. NotI digestion of pFAdV9-wt DNA: fragment sizes are 26 kb, 11.4 kb, 6 kb, 4 kb, and 1.4 kb; NotI digestion of lane .DELTA.0-2; fragment sizes are 26 kb; 11.4 kb, 5.9 k and 4 kb; Lane .DELTA.0-2 PCR product of 1619 pb and wt PCR product of 3107 pb; (C) Cytopathic effect (CPE) and expression of mCherry by RecFAdVs.

[0034] FIG. 4 shows the deletion of ORFs 1 and 2. (A) Flowchart of the generation of pFAdV9-.DELTA.1-2-RED: ORF1, 1A, 1B, 1C and 2 were replaced with the CAT cassette flanked on both sides with SwaI sites to generate pFAdV9-.DELTA.1-2CAT by homologous recombination. The CAT cassette was removed by SwaI digestion followed by religation to generate unmarked pFAdV9-.DELTA.1-2SwaI. The mCherry coding sequence was then cloned into SwaI site to generate pFAdV9-.DELTA.1-2RED, MLP, major late promoter; ITR, inverted terminal repeat; (B) and (D) NotI digestions and PCR amplification of DNA to verify the FAdmids and the corresponding viruses. Lanes .DELTA.1-2, pFAdV9-.DELTA.1-2RED or passage 3 virus; lanes wt: pFAdV9-wt or virus; M: 1 kb DNA ladder. NotI digestion of pFAdV9-wt DNA: fragment sizes are 26 kb, 11.4 kb, 6 kb, 4 kb, and 1.4 kb; NotI digestion of lane .DELTA.1-2; fragment sizes are 26 kb; 11.4 kb, 6.2 k and 4 kb; Lane .DELTA.1-2 PCR product of 1912 pb and wt PCR product of 3107 pb; (C) Expression of mCherry by RecFAdVs.

[0035] FIG. 5 shows the right end sequence of FAdV-9. Function of TR-2, ORF17 and ORF11 is unknown; TR-2: the longest repeat region is composed of 13 contiguous 135-bp-long direct repeats; ORF17 and 11 are probably membrane glycoproteins.

[0036] FIG. 6 shows the deletion of ORF17. (A) Flow chart of the generation of pFAdV9-.DELTA.17. ORF17 was replaced with the CAT cassette flanked on both sides with SwaI sites by homologous recombination to generate pFAdV9-.DELTA.17 (B) and (C) NotI digestions and PCR amplification of DNA to verify the FAdmids and the corresponding viruses. Lanes .DELTA.17, pFAdV9-.DELTA.17 or passage 3 virus; lanes wt: pFAdV9-wt or virus; M: 1 kb DNA ladder. NotI digestion of pFAdV9-wt DNA: fragment sizes are 26 kb, 11.4 kb, 6 kb, 4 kb, and 1.4 kb; NotI digestion of lane .DELTA.17; fragment sizes are 26,381 pb; 11,485 pb, 6056 pb, 4078 pb and 1391 pb; Lane .DELTA.17 PCR product of 2822 pb and wt PCR no product.

[0037] FIG. 7 shows the deletion of TR2, ORFs 17 and 11. (A) Flowchart of the generation of pFAdV9-.DELTA.TR2-17-11EGFP: TR2, ORF17, ORF11 were replaced with the CAT cassette flanked on both sides with SwaI sites to generate pFAdV9-.DELTA.TR2-17-11CAT by homologous recombination. The CAT cassette was removed by SwaI digestion followed by religation to generate unmarked pFAdV9-.DELTA.TR2-17-11SwaI. The EGFP cassette was then cloned into the SwaI site to generate pFAdV9-.DELTA.TR2-17-11EGFP, MLP, major late promoter; ITR, inverted terminal repeat; EGFP cassette, CMV-EGFP, (B) and (D) NotI digestions and PCR amplification of DNA to verify the FAdmids and the corresponding viruses. Lanes .DELTA.TR2; pFAdV9-.DELTA.TR2-17-11EGFP or passage 3 virus; lanes wt: pFAdV9-wt or virus; M: 1 kb DNA ladder. NotI digestion of pFAdV9-wt DNA: fragment sizes are 26 kb, 11.4 kb, 6 kb, 4 kb, and 1.4 kb; NotI digestion of lane .DELTA.TR2; fragment sizes are 21 kb, 11.4 kb, 6 kb, 4 kb, 3 kb and 1.4 kb; Lane .DELTA.TR2 PCR product of 3014 pb and wt PCR product of 4966 pb; (C) Expression of mCherry by RecFAdVs.

[0038] FIG. 8 shows that the recombinant FAdV-9 viral vector is a dual delivery vector capable of expressing exogenous nucleotide sequences at the left end and right end. (A) Flowchart of the generation of pFAdV9-.DELTA.1-2RED/TR2-17-11EGFP. TR2, ORF17 and ORF11 of pFAdV9-.DELTA.1-2RED were replaced with CAT cassette flanked on both sides with SwaI sites to generate pFAdV9-.DELTA.1-2RED.DELTA.TR2-17-11CAT by homologous recombination. The CAT cassette was replaced by EGFP cassette by SwaI digestion followed by ligation to generate pFAdV9-.DELTA.1-2RED/TR2-17-11EGFP. MLP, major late promoter; ITR, inverted terminal repeat; EGFP cassette, CMV-EGFP, (B) and (D) NotI digestions and PCR amplification of DNA to verify the FAdmids and the corresponding viruses. Lanes .DELTA.Dual: pFAdV9-.DELTA.1-2RED/TR2-17-11EGFP or passage 3 viruses. Lane M: 1 kb DNA ladder. Lanes wt: pFAdV9-wt or virus. NotI digestion of pFAdV9-wt DNA: the fragment sizes are 26 kb; 11.4 kb, 6 kb, 4 kb and 1.4 Kb; Lanes .DELTA.Dual NotI digestion: the fragments sizes are 21 kb, 11.4 kb, 6.2 kb, 4 kb and 3 kb; Lane .DELTA.Dual left PCR product of 1912 pb and wt PCR product of 3017 pb; Lane .DELTA.Dual right PCR product of 3014 pb and wt PCR product of 4966 pb.

[0039] FIG. 9 shows FAdV-9 based rec viruses with reporter genes.

[0040] FIG. 10(A) shows the nucleotide sequence of one embodiment of a recombinant FAdV-9 viral vector as described herein wherein, ORF1, ORF2, TR2, ORF17 and ORF11 have been deleted; (B) nucleotide sequences of genes inserted into recombinant viruses: Enhanced green fluorescent protein (EGFP, SEQ ID NO: 2) and mCherry red (SEQ ID NO: 3); (C) nucleotide sequences of promoters inserted into recombinant genes: Human cytomegalovirus immediate early promoter (CMV; SEQ ID NO: 4), CMV enhancer/chicken .beta.-actin promoter (CAG, SEQ ID NO: 5), Human elongation factor 1 alpha promoter (EF1.alpha., SEQ ID NO: 6), L2R promoter (SEQ ID NO: 7) and .beta.-actin promoter (SEQ ID NO: 8); and (D) nucleotide sequence of enhancer/regulatory used in recombinant viruses: Woodchuck hepatitis virus post-transcriptional regulatory element (WPER, SEQ ID NO: 9).

[0041] FIG. 11 shows a schematic representation of pCI-Neo. The plasmid pCI-Neo (Promega) was chosen to create dual expression constructs due to its' unique restriction enzyme sites in and before the multiple cloning site, as well as the neomycin resistance gene.

[0042] FIG. 12 shows generation of recombinant FAdV-9.DELTA.4 viruses. (A) The EGFP expression cassette was amplified by PCR from an intermediate pHMR construct, or gel extracted after double digestion with BamHI and BglII. (B) pFAdV-9.DELTA.4 was linearized and digested with SwaI. (C) Both the EGFP cassette and the linearized pFAdV-9.DELTA.4 was co-transformed into E. coli BJ5183 cells to undergo homologous recombination. (D) The resulting plasmid was transformed into E. coli DH5.alpha. cells, propagated, screened by NotI digestion, and eventually linearized with PacI to release the viral genome from the plasmid. The linear recombinant viral DNA was transfected into CH-SAH cells. (E) Recombinant virus was collected in the supernatent. This procedure was carried out for each EGFP expression cassette, resulting in the viruses: FAdV-9.DELTA.4-CMV-EGFP, FAdV-9.DELTA.4-CMV-EGFP-WPRE, FAdV-9.DELTA.4-CAG-EGFP, FAdV-9.DELTA.4-CAG-EGFP-WPRE, FAdV-9.DELTA.4-EF1.alpha.-EGFP, and FAdV-9.DELTA.4-EF1.alpha.-EGFP-WPRE.

[0043] FIG. 13 shows a schematic representation of EGFP/luciferase dual-expression plasmids. (A) EGFP was PCR amplified from pEGFP-N1 and cloned into the multiple cloning site of pCI-Neo. The neomycin resistance (NeoR) gene from pCI-Neo was removed by restriction enzyme digestion. Firefly luciferase was PCR amplified from pGL-4.17 and cloned under the SV40 promoter, resulting in a dual-expression plasmid. (B) The plasmid pCMV-EGFP-Luc was digested with SpeI and EcoRI to clone in promoters of interest, resulting in five vectors controlling EGFP under different promoters and luciferase under the SV40 promoter: pCMV-EGFP-Luc, pCAG-EGFP-Luc, pEF1.alpha.-EGFP-Luc, p.beta.-actin-EGFP-Luc, and pL2R-EGFP-Luc. Five additional plasmids were made by non-directionally cloning the Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) into each dual-expression plasmid at the NotI site, resulting in the constructs: pCMV-EGFP-WPRE-Luc, pCAG-EGFP-WPRE-Luc, pEF1.alpha.-EGFP-WPRE-Luc, p.beta.-actin-EGFP-WPRE-Luc, and pL2R-EGFP-WPRE-Luc.

[0044] FIG. 14 shows a comparison of EGFP expression with fluorescence microscopy. CH-SAH cells were seeded in 35 mm plates (1.8.times.10.sup.6 cells/dish) and transfected with 2 .mu.g of plasmid DNA. Fluorescence of EGFP was measured by microscopy at 12, 24, 36, 48, 60, and 72 hours post-transfection. A mock transfection was used as the negative control while pEGFP-N1 was the positive control.

[0045] FIG. 15 shows normalized EGFP expression over-time. CH-SAH cells were seeded in 35 mm plates (1.8.times.10.sup.6 cells/plate) and transfected with 2 .mu.g of plasmid DNA. At each time-point, transfected cells were washed, trypsinized, and resuspended in PBS. After three freeze-thaw cycles, samples were centrifuged and the protein concentration of the collected supernatant was measured using a nanodrop. Fluorescence of EGFP was measured in a microplate reader at 480 and 528 nm excitation and emission wavelengths, respectively. Luminescence of luciferase was measured using a Pierce Firefly Luciferase Glow Assay kit (Thermo). The dual-expression (fluorescence/luminescence) of each construct was normalized to the expression level of pCMV-EGFP-Luc (normalized to 1.0) at each time-point. The t-test was used to determine the significance of expression compared to pCMV-EGFP-Luc, indicated by an asterisks (*), where P<0.05.

[0046] FIG. 16 shows a schematic representation of intermediate constructs used to generate recFAdVs. (A) FAdV-9 DNA flanking the .DELTA.4 deletion site (VF1 and VF2) was PCR amplified and directionally cloned into the dual-expression plasmid pCMV-EGFP-Luc. The resulting construct, pHMR-CMV-EGFP, was used downstream to generate the recombinant virus FAdV-9.DELTA.4-CMV-EGFP. (B) Five additional intermediate constructs were generated, resulting in the plasmids: pHMR-CAG-EGFP, pHMR-EF1.alpha.-EGFP, pHMR-CMV-EGFP-WPRE, pHMR-CAG-EGFP-WPRE, and pHMR-EF1.alpha.-EGFP-WPRE.

[0047] FIG. 17 shows agarose gel electrophoresis screen of NotI digested FAdmids. FAdmids derived from homologous recombination between pFAdV-9.DELTA.4 and pHMR plasmids were digested with NotI and the resulting banding patterns were screened on agarose gel. Digested pFAdV-9.DELTA.4 has an expected banding pattern of 4 kb, 5.1 kb, 13.7 kb, and 23.8 kb. Recombinant FAdmids with an EGFP expression cassette contain an additional NotI site for screening. Diagnostic bands corresponding to each cassette are as follows: CMV-EGFP=2.2 kb, CMV-EGFP-WPRE=2.2 kb and 0.55 kb, CAG-EGFP=2.9 kb, CAG-EGFP-WPRE=0.55 kb, EF1.alpha.-EGFP=2.7 kb, and EF1.alpha.-EGFP-WPRE=2.7 kb and 0.55 kb. White arrows point to the diagnostic bands.

[0048] FIG. 18 shows viral growth curves. One-step growth curves for each recombinant fowl adenovirus were determined in CH-SAH cells. Cells were seeded in 35 mm plates (1.8.times.10.sup.6 cells/plate) and infected at an MOI of 5. Both intracellular and extracellular virus was collected between 0-72 h.p.i. One-step growth curves were repeated in duplicate for each sample and all extracellular virus was titered by plaque assay.

[0049] FIG. 19 shows cytopathic effect of recombinant FAdV matches FAdV-9.DELTA.4. CH-SAH cells were plated in 35 mm plates (1.8.times.10.sup.6 cells/plate) and infected at an MOI of 5. Cytopathic effect of recombinant viruses was compared to the "wild-type" control FAdV-9.DELTA.4. It was observed that all recombinant viruses (data not shown) had CPE similar to FAdV-9.DELTA.4, evidenced by cell rounding and detachment using a bright-field microscope. However, fluorescence of EGFP by recFAdVs, for example FAdV-9.DELTA.4-CAG-EGFP-WPRE, was observed using fluorescence microscopy.

[0050] FIG. 20 shows the time course of EGFP expression in CH-SAH cells. CH-SAH cells were seeded in 35 mm plates (1.8.times.10.sup.6 cells/plate) and infected with recFAdV-9s at an MOI of 5. At each time-point, infected cells were collected, washed, and resuspended in PBS. After three freeze-thaw cycles, samples were centrifuged and the protein concentration of the collected supernatant was measured with a nanodrop. The absolute fluorescence of EGFP from 50 .mu.g of whole cell lysate was measured with a microplate reader at 480 and 528 nm excitation and emission wavelengths, respectively. Uninfected (mock) or FAdV-9.DELTA.4 infected cells did not show any EGFP fluorescence.

[0051] FIG. 21 shows western immunoblot of EGFP production over-time. EGFP expression by recFAdV-9s in CH-SAH cells was compared over 48 hpi, along with FAdV-9.DELTA.4 (negative control), uninfected mock (negative control), and pEGFP-N1 transfected CH-SAH cells (positive control). CH-SAH cells were seeded in 35 mm plates (1.8.times.10.sup.6 cells/dish) and infected with recFAdV-9s at an MOI of 5. At each time-point whole cell lysates were collected and protein concentrations were determined by Bradford assay. Six .mu.g of each sample was loaded into two gels, one to be probed with anti-EGFP antibody (1:1,000) followed by a secondary anti-mouse HRP conjugated antibody (1:5,000), the other gel to be probed with anti-actin antibody (1:200) followed by a secondary anti-goat HRP conjugated antibody (1:20,000). All bands appeared at the expected size (EGFP=27 kDa and actin=42 kDa) as determined from a molecular weight marker (not shown).

[0052] FIG. 22 shows polyvalent recombinant FadV-9 with H5, H7 and HN.

DETAILED DESCRIPTION OF THE INVENTION

[0053] The inventors have determined that recombinant FAdV-9 with deletions on the left and/or right side of the genome are useful as viral vectors.

[0054] In one embodiment of the invention, the recombinant FAdV-9 viral vector has a deletion at the left end of the genome. In one embodiment, the deletion at the left end of genome comprises a deletion of one or more of ORF0, ORF1 and ORF2.

[0055] In one embodiment, the recombinant FAdV-9 viral vector has a deletion at the right end of the genome. In one embodiment, the deletion at the right end of the genome comprises a deletion of one or more of ORF19, TR2, ORF17 and ORF11.

[0056] In one additional embodiment, the recombinant FAdV-9 viral vector has deletions at both the left end and right end of the genome. In one embodiment, the FAdV-9 viral vector has a deletion of ORF0, ORF1, ORF2, TR2, ORF17 and ORF11. In one embodiment, the recombinant FAdV-9 viral vector has a deletion of ORF1, ORF2, TR2, ORF17 and ORF11. In one embodiment, the FAdV-9 viral vector has a deletion of ORF1, ORF2, and ORF 19. In one embodiment, the recombinant FAdV9-viral vector has a deletion of ORF1, ORF2 and ORF19, TR2 or ORF11.

[0057] As shown in the Examples, recombinant FAdV-9 viral vectors with deletions on the left side and right side, including deletion of TR2, are stable and may be used to drive transgene expression.

[0058] The FAdV viral vectors described herein provide a number of advantages. For example, in some embodiments the FAdV viral vectors allow for the production of mono and polyvalent vaccines. The value of having a vector capable of expressing dual or even multivalent antigens is that only one vaccine would be needed to protect against to ore more diseases. In one embodiment, the vectors are capable of incorporating large segments of foreign DNA, optionally up to 7.7 kb of foreign DNA. In one embodiment, the vectors are stable and safe for use in animals such as chickens. In one embodiment, viral vectors are easy to produce, and produce high viral titers. In one embodiment, there are a variety of serotypes of FAdVs. In one embodiment, the viral vectors have no pre-existing immunity in humans and may be used in human gene therapy.

[0059] In one embodiment, the FAdV-9 viral vectors described herein may include one or more exogenous nucleotide sequences (also referred to herein as transgenes).

[0060] In an embodiment of the invention, the exogenous nucleotide sequence is selected from antigenic sequences against influenza, infectious laryngotracheitis, infectious bronchitis, bursa of Fabricius' infection (Gumboro), hepatitis, viral rhinotracheitis, infectious coryza, Mycoplasma hyopneumonieae, pasteurellosis, Porcine Respiratory and Reproductive Syndrome (PRRS), circovirus, bordetellosis, parainfluenza, or any other antigen which size allows its insertion into the corresponding viral vector.

[0061] In another embodiment, the exogenous nucleotide sequence is selected from antigenic sequences against Avian influenza, Laryngotracheitis (LT), Newcastle disease (NDV), infectious anemia, Inclusion bodies, Infectious Bronchitis (IB), Metapneumovirus (MPV) or Gumboro.

[0062] In one preferred embodiment of the invention, the exogenous nucleotide sequence comprises, consists essentially of or consists of a sequence corresponding to at least one gene disclosed in Table 1, or a homolog thereof. As used herein, the term "homolog" of a gene is intended to denote a gene with at least 85%, of at least 90%, at least 98% or at least 99% sequence identity to the gene, and having a biological activity of the same nature. In one embodiment, the vector described herein comprises 2, 3 or 4 sequences corresponding to the genes disclosed in Table 1, or a homolog thereof. Optionally, the exogenous sequences may be inserted into the vector at a single insertion site or multiple different insertions sites.

TABLE-US-00001 TABLE 1 DISEASE GENE GEN BANK ACCESSION Avian influenza Mexican Avian influenza H5N2 FJ864690.1 (SEQ ID NO: 10) hemagglutinin, strain 435 Avian influenza H7N3 JX397993.1 (SEQ ID NO: 11) hemagglutinin Jalisco 2012 Avian influenza H7N3 KR821169 (SEQ ID NO: 12) hemagglutinin Guanajuato 2015 Laryngotracheitis (LT) Laryngotracheitis glycoprotein B, EU104965.1 (SEQ ID NO: strain USDA 13) Laryngotracheitis glycoprotein D, JN542534.1|: 132317-133621 strain USDA (SEQ ID NO: 14) Newcastle (NDV) HN gene strain LaSota |KC844235.1|: 6412-8145 (SE ID NO: 15) HN gene strain Newcastle disease KF910962.1 (SEQ ID NO: 16) virus isolate Chicken/M08- 3313/Mexico/2008 infectious anemia VP1 gene from strain Chicken KJ872513.1|: 832-2181 (SEQ anemia virus isolate CAV-10 ID NO: 17) VP2 gene from strain Chicken AIV09094.1 (SEQ ID NO: 18) anemia virus isolate CAV-10 Inclusion bodies Fowl adenovir us 4 short fiber AY340863.1 (SEQ ID NO: 19) gene Infectious Bronchitis Infectious bronchitis virus strain EU359651.1 (SEQ ID NO: (IB) ArkDPI-derived commercial 20) vaccine D spike glycoprotein gene Metapneumovirus Avian metapneumovirus isolate |JF424833.1|: 2943-45599 (MPV) IT/Ty/A/259-01/03, F gene (SEQ ID NO: 21) Gumboro Gumboro VP2

[0063] The FadV-9 viral vectors described herein can be prepared using recombinant technologies such as PCR amplification of a nucleotide sequence of interest, by identifying the antigenic sites from an isolation of the origin-pathogen, to be further inserted, amplified in the viral vector. The insertion may be made using standard molecular biology techniques, such as restriction enzymes and DNA ligases, amongst others. The infectious clone thus produced is introduced into a suitable cell line for the production of the recombinant virus. For example, in one embodiment, the methodologies required for the construction of a FAdV-9 viral vectors are described in the present Examples and the procedures described for the construction of the FAdV-9 infectious clone (FAdmid) (Ojkic, D. & Nagy, E. (2001), The long repeat region is dispensable for fowl adenovirus replication in vitro. Virology 283, 197-206). This procedure utilizes homologous recombination between the viral genomic DNA and the linearized plasmid containing both ends of the genome flanking a backbone vector (pWE-Amp with PacI sites introduced). Next, a foreign gene of interest with a promoter to drive its expression is inserted in suitable genomic regions of the infectious clone that are dispensable or non-essential (Corredor and Nagy, 2010a and 2010b), and introduced into a suitable cell line.

[0064] The viral vector of the present invention can be used, for example, for the preparation and administration of immunogenic compositions comprising at least the viral vector as described herein and an exogenous nucleotide sequence coding for at least one antigenic site of a disease of concern inserted therein. Optionally, the viral vector of the present invention is a dual delivery vector that can be used to drive the expression of two or more exogenous nucleotide sequences. In one embodiment, the two or more exogenous nucleotide sequences are under the control of different promoters. In one embodiment the promoters are selected from the group consisting of CMV, CAG, EF1.alpha., .beta.-actin and L2R.

[0065] In one embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein ORF0, ORF1, ORF2, TR2, ORF17 and ORF11 have been deleted. SEQ ID NO: 1 corresponds to the complete genome sequence of Fowl adenovirus D (Genbank accession no. AC_000013.1). In one embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of ORF0, ORF1, ORF2, TR2, ORF17 and ORF11 have been deleted. In a preferred embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all of part of ORF1, ORF2, TR2, ORF17 and ORF11 have been deleted. In one embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein ORF1, ORF2, TR2, ORF17 and ORF11 have been deleted.

[0066] In another embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of ORF1, ORF2 and ORF19 have been deleted. In one embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of ORF1, ORF2, and ORF19 have been deleted.

[0067] In another embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of ORF1, ORF2 and ORF19, TR2 or ORF11 have been deleted. In one embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of ORF1, ORF2 and ORF19, TR2 or ORF11 have been deleted.

[0068] In another embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of nucleotides 575 to 2753 have been deleted. This sequence (nucleotides 575 to 2753) includes ORF0, ORF1A, ORF1B, ORF1C and ORF2. In another embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 575 to 2753 have been deleted.ln another embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of nucleotides 847 to 2753 have been deleted. This nucleotide sequence (nucleotide 847 to 2753) includes ORF1A, ORF1B, ORF1C and ORF2. In another embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of nucleotides 847 to 2753 have been deleted.

[0069] In another embodiment, the fowl adenovirus described herein comprises a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of nucleotides 38,807 to 42,398 have been deleted. This nucleotide sequence (nucleotides 38,807 to 42,398) includes TR-2, ORF17 and ORF11. In another embodiment, the FAdV-9 viral vector described herein comprises or consists of a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the sequence shown in SEQ ID NO: 1, wherein all or part of nucleotides 38,807 to 42,398 have been deleted.

[0070] In one embodiment, the viral vector comprises or consists of a nucleotide sequence with sequence identity to a sequence comprising or consisting of SEQ ID NO: 1, wherein at least 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000 nucleotides corresponding to all or part of nucleotides 575 to 2753 and/or 38,807 to 42,398 of SEQ ID NO: 1 have been deleted.

[0071] In one embodiment, the viral vector comprises or consists of a nucleotide sequence with sequence identity to a sequence comprising or consisting of SEQ ID NO: 1, wherein at least 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000 nucleotides corresponding to all or part of nucleotides 847 to 2753 and/or 38,807 to 42,398 of SEQ ID NO: 1 have been deleted.

[0072] In another embodiment, the viral vector comprises or consists of a nucleotide sequence with sequence identity to a sequence comprising or consisting of SEQ ID NO: 1, wherein at least 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000 nucleotides corresponding to all or part of nucleotides 847 to 2753 and 34,220 to 36,443 have been deleted.

[0073] In another embodiment, the viral vector comprises or consists of a nucleotide sequence with sequence identity to a sequence comprising or consisting of SEQ ID NO: 1, wherein at least 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500 or 5000 nucleotides corresponding to all or part of nucleotides 847 to 2753, 34,220 to 36,443, and 38,807 to 40,561 or 41,461 to 42398 have been deleted.

[0074] At least one exogenous nucleotide sequence is optionally inserted into the FAdV-9 viral vector described herein. Accordingly, in one embodiment, the FAdV-9 nucleotide sequence with the deletions described herein is not present in the vector as one contiguous sequence but rather includes sections of contiguous sequences interrupted by at least one, and optionally at least two, three or four, exogenous nucleotide sequences. Therefore, in one embodiment, the fowl adenovirus described herein comprises one or more nucleotide sequence(s) with sequence identity to the one or more sequences shown in SEQ ID NO: 1, wherein at least one of ORF0, ORF1, ORF2, TR2, ORF17 and ORF11 have been deleted, and the nucleotide sequence comprises at least two, three, four or five contiguous sequences.

[0075] The number of nucleotides deleted from the FAdV-9 varies. In one embodiment, 1000 to 7000 nucleotides are deleted, optionally split between the left and the right end of the genome. In other embodiments, 1500 to 6000 nucleotides are deleted. In one embodiment, at least 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 5000 or 6000 nucleotides are deleted.

[0076] The size of the exogenous nucleotide sequence(s) inserted into the viral vector described herein also varies. In one embodiment, based on the 105% adenovirus stability rule, the capacity of the vector is up to 7751 bp. In other embodiments, 1000 to 7000 nucleotides are inserted, optionally split between the left and the right end of the genome. For example, one foreign gene may be inserted into the left end and a second foreign gene may be inserted into the other end. In other embodiments, 1500 to 6000 nucleotides are inserted. In one embodiment, at least 1000, 2000, 3000, 4000, 5000 or 6000 exogenous nucleotides are inserted.

[0077] In one embodiment, the FAdV-9 viral vector has a deletion at the left end of the genome of about 2291 base pairs, optionally between about 1900 and 2500 base pairs. In one embodiment, the FAdV-9 viral vector has a deletion at the right end of the genome of about 3591 base pairs. In one embodiment, the FAdV-9 viral vector has a deletion at the right end of the genome of between about 3000 base pairs and 4000 base pairs.

[0078] Sequence identity is typically assessed by the BLAST version 2.1 program advanced search (standard default parameters; Altschul, S. F., Gish, W., Miller, W., Myers, E. W. & Lipman, D. J. (1990) "Basic local alignment search tool." J. Mol. Biol. 215:403_410). BLAST is a series of programs that are available online through the U.S. National Center for Biotechnology Information (National Library of Medicine Building 38A Bethesda, Md. 20894) The advanced Blast search is set to default parameters. References for the Blast Programs include: Altschul, S. F., Gish, W., Miller, W., Myers, E. W. & Lipman, D. J. (1990) "Basic local alignment search tool." J. Mol. Biol. 215:403-410; Gish, W. & States, D. J. (1993) "Identification of protein coding regions by database similarity search." Nature Genet. 3:266-272.; Madden, T. L., Tatusov, R. L. & Zhang, J. (1996) "Applications of network BLAST server" Meth. Enzymol. 266:131-141; Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. (1997) "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs." Nucleic Acids Res. 25:3389-3402); Zhang, J. & Madden, T. L. (1997) "PowerBLAST: A new network BLAST application for interactive or automated sequence analysis and annotation." Genome Res. 7:649-656).

[0079] As used herein, "viral vector" refers to a recombinant adenovirus that is capable of delivering an exogenous nucleotide sequence into a host cell. For example, in one embodiment, the viral vector comprises restriction sites that are suitable for inserting an exogenous nucleotide sequence into the vector. In one embodiment, one or more nucleotide sequences which are not required for the replication or transmission of FAdV-9 described herein are deleted in the nucleotide sequence of the viral vector. For example, in one embodiment nucleotide sequences at the left and/or right end of the FAdV-9 genome are deleted in the recombinant FAdV-9 viral vector. In one embodiment, nucleotide sequences corresponding to one or more of ORFs 0-2, ORF19, TR2, ORF17 and ORF11 are deleted in the recombinant FAdV-9 viral vector. In one embodiment, the viral vector includes one or more exogenous control sequences such as promoters or cloning sites useful for driving the expression of transgenes. In one embodiment the promoters are selected from the group consisting of CMV (SEQ ID NO: 4), CAG (SEQ ID NO: 5), EF1.alpha. (SEQ ID NO: 6), .beta.-actin (SEQ ID NO: 8) and L2R (SEQ ID NO: 7).

[0080] In one embodiment, the viral vector comprises an exogenous nucleotide sequence coding for a polypeptide of interest. In one embodiment, the polypeptide of interest is an antigen from a disease of concern. For example, in one embodiment, the viral vector comprises an exogenous nucleotide sequence coding for at least one antigenic site of a disease of concern. Exogenous nucleotide sequences coding for a polypeptide of interest can readily be obtained by methods known in the art such as by chemical synthesis, screening appropriate libraries or by recovering a gene sequence by polymerase chain reaction (PCR).

[0081] With respect to the present disclosure, diseases of concern include, but are not limited to, influenza, infectious laryngotracheitis (ILT), infectious bronchitis (IB), infectious bursal disease (Gumboro), hepatitis, viral rhinotracheitis, infectious coryza, Mycoplasma hyopneumonieae, pasteurellosis, Porcine Respiratory and Reproductive Syndrome (PRRS), circovirus, bordetellosis, parainfluenza, Avian influenza, Newcastle disease (NDV), infectious anemia, Inclusion bodies hepatitis (IBH), and Metapneumovirus (MPV).

[0082] In one embodiment, the viral vector is adapted to express an exogenous nucleotide sequence in a host cell. For example, in one embodiment the viral vector comprises control sequences capable of affecting the expression of an exogenous nucleotide sequence in a host. For example, the viral vectors described herein may include one or more control sequences such as a transcriptional promoter, an enhancer, an optional operator sequence to control transcription, a sequence encoding suitable mRNA ribosomal binding sites, alternative splicing sites, translational sequences, or sequences which control the termination of transcription and translation. Optionally, the viral vector comprises different control sequences at the left end and right end of the vectors. In one embodiment the promoters are selected from the group consisting of CMV (SEQ ID NO: 4), CAG (SEQ ID NO: 5), EF1.alpha. (SEQ ID NO: 6), .beta.-actin (SEQ ID NO: 8) and L2R (SEQ ID NO: 7) and the enhancer may be WPRE (SEQ ID NO: 9).

[0083] In one embodiment, the viral vector comprises one or more exogenous nucleotide sequences operably linked to one or more control sequences. In one embodiment, the viral vector comprises an insertion site adjacent to one or more control sequences such that when an exogenous nucleotide sequence is inserted into the vector, the exogenous nucleotide sequence is operably linked to the control sequences.

[0084] As used herein, nucleotide sequences are "operably linked" when they are functionally related to each other. For example, a promoter is operably linked to a coding sequence if it controls the transcription of the sequence; a ribosome binding site is operably linked to a coding sequence if it is positioned so as to permit translation. Optionally, sequences that are operably linked are contiguous sequences in the viral vector.

[0085] In one embodiment, the viral vector described herein includes a sequence suitable for the biological selection of hosts containing the viral vector such as a positive or negative selection gene.

[0086] Other methods known in the art, such as recombinant technologies including but not limited to those disclose in disclosed by Sambrook et al (Sambrook J et al. 2000. Molecular Cloning: A Laboratory Manual (Third Edition), Cold Spring Harbor Laboratory Press), are also suitable for preparing the nucleotide sequences and viral vectors as described herein.

[0087] Optionally, the viral vectors and methods described herein may be used for gene therapy in animal subjects in need thereof. For example, in one embodiment, the viral vectors described herein may be used for the delivery and expression of a therapeutic nucleotide sequence or nucleotide encoding a therapeutic protein. In one embodiment, there is provided a method of gene therapy comprising administering to a subject in need thereof a viral vector or composition as described herein, wherein the viral vector comprises an exogenous nucleotide sequence encoding a therapeutic nucleotide sequence or protein.

[0088] Another aspect of the present disclosure includes an immunogenic composition comprising a recombinant FAdV-9 viral vector as described herein. In one embodiment, the immunogenic compositions can be prepared by known methods for the preparation of compositions for the administration to animals including, but not limited to, humans, livestock, poultry and/or fish. In one embodiment, an effective quantity of the viral vector described herein is combined in a mixture with a pharmaceutically acceptable carrier. Suitable carriers are described, for example in Remington's Pharmaceutical Sciences (Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., USA) or Handbook of Pharmaceutical Additives (compiled by Michael and Irene Ash, Gower Publishing Limited, Aldershot, England (1995). On this basis, the compositions include, albeit not exclusively, solutions of the viral vectors describes herein in association with one or more pharmaceutically acceptable carriers or diluents, and may be contained in buffered solutions with a suitable pH and/or be iso-osmotic with physiological fluids. In one embodiment, the immunogenic composition comprises an adjuvant.

[0089] The novel FAdV-9 viral vectors, associated methods and uses will be more clearly illustrated by means of the following description of specific examples.

EXAMPLE 1

Materials and Methods

1.1 Cell Culture and Viruses

[0090] Chicken hepatoma cells (CH-SAH cell line) were maintained in Dulbecco's Modified Eagle's Medium/Nutrient Mixture F-12 Ham (DMEM-F12) (Sigma) plus 200 mM L-glutamine and 100 U/ml penicillin-streptomycin (PenStrep, Sigma) with 10% non-heat inactivated fetal bovine serum (FBS) as described (Alexander, H. S., Huber, P., Cao, J., Krell, P. J., Nagy, E. 1998. Growth Characteristics of Fowl Adenovirus Type 8 in a Chicken Hepatoma Cell Line. J. Virol. Methods. 74, 9-14.). Recombinant FAdVs were generated using the FAdV-9.DELTA.4 deletion virus described by Corredor and Nagy (2010b) as the base. Propagation of all viruses were carried out in CH-SAH cells as described by Alexander et al. (1998).

1.2 General DNA Manipulation

1.2.1 Bacterial Cultures and Plasmid Isolation

[0091] Escherichia coli DH5.alpha. cells were the bacterial host for all plasmids described, while E. coli BJ5183 cells were used for homologous recombination to generate recombinant FAdmids. Bacterial cultures were grown on selective Luria-Bertani (LB) liquid or agar (16 mg/ml) growth medium containing ampicillin (100 .mu.g/ml) at 37.degree. C. Single E. coli colonies were picked, inoculated in 5 ml of LB medium supplemented with 0.1% ampicillin to select for growth of bacteria containing a transformed ampicillin resistant plasmid. Incubation and growth was for approximately 16 hours. Chemically (CaCl.sub.2) competent DH5.alpha. E. coli were prepared and transformed with either 10 .mu.l of ligation product or 1 .mu.l purified plasmid DNA (Sambrook, J., Russel, D. W. 2001. Molecular Cloning: A Laboratory Manual, Volume 1. 3.sup.rd Ed. Cold Spring Harbor Laboratory Press, Cold Spring, N.Y., USA). All plasmids were isolated using either the EZ-10 Spin Column Plasmid DNA Mini-prep kit (Bio Basic) or the PureLink HiPure Plasmid Midiprep kit (Invitrogen) (for plasmids larger than 40 kb in size or when a high concentration of DNA was needed) as per the manufacturer's protocol.

1.2.2 PCR and Restriction Enzyme Digestion

[0092] DNA was amplified by polymerase chain reaction (PCR) during the cloning of all dual-expression constructs and recombinant viruses. For all cloning, PCR amplification was conducted using a Kod Hot Start Polymerase kit (Novagen). However, Taq polymerase was used when screening a plasmid by PCR. Unless stated otherwise, the PCR conditions for both Kod and Taq polymerases are summarized in Table 2. All PCR reactions were carried out with a Mastercycler Pro (Eppendorf).

[0093] Restriction enzyme (RE) digestions were carried out for both Fast Digest enzymes (Fermentas) and enzymes from New England BioLabs (NEB). All reactions occurred as per the manufacturer's protocol (per enzyme). Digestion reactions were always performed at 37.degree. C., and samples were heat inactivated in a Mastercycler Pro (Eppendorf) thermocycler.

[0094] Unless otherwise stated, all DNA samples were subjected to electrophoresis at 100V in 0.8% agarose gels containing 1.times. RedSafe.TM. (iNtRON Biotechnology). 6.times. DNA loading buffer [0.25% (w/v) bromophenol blue, 40% sucrose (w/v) in water] and 1.times. Tris-acetate-EDTA (TAE) buffer were used for electrophoresis of DNA samples.

1.2.3 Plasmid Construction and Transformation

[0095] After RE digestion and gel purification, PCR amplified DNA and plasmid were ligated together with T4 DNA ligase (Invitrogen). Unless stated otherwise, ligations were performed at a molar ratio of 1:1 insert to vector, overnight at 16.degree. C. Fifty .mu.l of CaCl.sub.2 competent DH5.alpha. E. coli were mixed with 10 .mu.l of ligation product or 1 .mu.l purified plasmid DNA. Competent cells and DNA were incubated on ice for 30 min, then heat shocked at 42.degree. C. for 1 min. Cells were recovered on ice for 3 min and 500 .mu.l of super optimal broth with catabolite repression (SOC) medium was added to each microcentrifuge tube. The cells were incubated at 37.degree. C. for 1 hr with agitation, then centrifuged and resuspended in 100 .mu.l of LB broth. The entire volume was spread on a LB agar plate containing ampicillin.

[0096] Homologous recombination occurred in E. coli BJ5183 cells. Two .mu.g of both promoter cassettes and linear FAdV-9.DELTA.4 DNA were mixed in 100 .mu.l of chemically competent BJ5183 cells. Mixtures were left on ice for 15 min, followed by a heat shock at 42.degree. C. for 1 min. Cells were recovered on ice for 20 min, and 1 ml of SOC medium was added to each microcentrifuge tube and transferred into a 5 ml glass culture tube. Cells were incubated for 2 hrs at 37.degree. C. with agitation, then centrifuged and resuspended in 100 .mu.l of LB broth. The entire volume of cells was spread onto a LB agar plate containing ampicillin.

1.2.4 Sequencing

[0097] All PCR products and plasmids were purified and sequenced by the ABI 3730 DNA sequencer (Laboratory Services Division, Guelph, ON). Sequence data were analysed using SnapGene Viewer (GSL Biotech).

1.3 Analysis of Promoter Expression

1.3.1 Generation of Dual-Expression Plasmid Constructs

[0098] The activity of five promoters (CMV, CAG, EF1.alpha., .beta.-actin, and L2R) and one enhancer element (WPRE) were compared by measuring the expression of EGFP compared to firefly luciferase under the SV40 promoter in transfected CH-SAH cells. The plasmids, pCI-Neo (Promega), pCAG-Puro, and pEF1.alpha.-Puro, were provided by Dr. Sarah Wootton (University of Guelph). Dual-expression plasmids were generated using the plasmid pCI-Neo as a backbone (FIG. 11), which contained the CMV promoter along with numerous unique RE sites. Both the CAG and EF1.alpha. promoter were sub-cloned from pCAG-Puro and pEF1.alpha.-Puro, respectively, into pCI-Neo using SpeI and EcoRI. The presence of each promoter was confirmed by sequencing using the primer pCI-Neo-F (Table 3). EGFP was amplified by PCR from pEGFP-N1 (Clontech Laboratories, Inc) with primers EGFP-F and EGFP-R (Table 3) at an annealing temperature of 60.degree. C. The resulting PCR product was gel extracted using the Wizard Plus SV Miniprep DNA Purification Kit (Promega). Both EGFP PCR product and pCI-Neo based plasmids (containing the CMV, CAG, or EF1.alpha. promoter) were subjected to double digestion with EcoRI and NotI for 1 hr at 37.degree. C. Both digested plasmid and PCR product were then separated in a gel and extracted using Wizard Plus SV Miniprep DNA Purification Kit (Promega), and ligated overnight at 4.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were PCR screened for the presence of the EGFP fragment with primers EGFP-F and EGFP-R. All positive colonies were confirmed by sequencing with EGFP-I-F primer (Table 3), resulting in the plasmids pCMV-EGFP, pCAG-EGFP, and pEF1.alpha.-EGFP.

[0099] The .beta.-actin promoter was PCR amplified from pCAG-Puro using the primers .beta.actin-F and .beta.actin-R (Table 3) with an annealing temperature of 60.degree. C. The resulting PCR product was gel extracted with the Wizard Plus SV Miniprep DNA Purification Kit (Promega). Both .beta.actin PCR product and pCAG-EGFP were subjected to double digestion with EcoRI and NotI for 1 hr at 37.degree. C. Digested plasmid and PCR product were then separated in a gel and extracted using Wizard Plus SV Miniprep DNA Purification Kit (Promega), and ligated overnight at 4.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were screened with RE for the presence of .beta.actin. All positive colonies were confirmed by sequencing using both pCI-Neo-F and EGFP-I-F primers (Table 3), resulting in the plasmid ppactin-EGFP.

[0100] The fowlpox virus L2R promoter was PCR amplified from pE68 (Zantinge, J. L., Krell, P. J., Derbyshire, J. B., Nagy, E. 1996. Partial transcriptional mapping of the fowlpox virus genome and analysis of the EcoRI L fragment. J. Gen. Virol. 77(4), 603-614) with the primers L2R-F and L2R-R (Table 3) at an annealing temperature of 60.degree. C. The resulting PCR product was gel extracted using the Wizard Plus SV Miniprep DNA Purification Kit (Promega). Both L2R PCR product and pCMV-EGFP were subjected to double digestion with EcoRI and NotI for 1 hr at 37.degree. C. Both digested plasmid and PCR product were then separated in a gel and extracted using Wizard Plus SV Miniprep DNA Purification Kit (Promega), and ligated overnight at 4.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were screened with RE for the presence of L2R. All positive colonies were confirmed by sequencing with both pCI-Neo-F and EGFP-I-F primers (Table 3), resulting in the recovery of the plasmid pL2R-EGFP.

[0101] Firefly luciferase was PCR amplified from pGL4.17 (Promega) with primers Luc-F and Luc-R (Table 3) at an annealing temperature of 55.degree. C. The resulting PCR product (1.6 kb) was gel extracted using the Wizard Plus SV Miniprep DNA Purification Kit (Promega). Both luciferase PCR product and promoter plasmids were subjected to double digestion with AvrII and BstBI for 1 hr at 37.degree. C. Digested plasmid and PCR product were then separated in a gel, removing the neomycin resistance (NeoR) cassette from each plasmid, and extracted using Wizard Plus SV Miniprep DNA Purification Kit (Promega), and ligated overnight at 4.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were PCR screened for the presence of luciferase. All positive colonies were confirmed by sequencing using SV40-F primer (Table 3).

[0102] Five additional dual-expression plasmids were generated to include the enhancer element WPRE. The WPRE element was PCR amplified from pWPRE (Dr. Sarah Wootton, University of Guelph) using the primers WPRE-F and WPRE-R (Table 2.2) with an annealing temperature of 55.degree. C. The PCR product was gel extracted using the Wizard Plus SV Miniprep DNA Purification Kit (Promega). Both WPRE PCR product and promoter were subjected to digestion with NotI for 1 hr at 37.degree. C. After digestion, plasmids were treated with alkaline phosphatase (calf intestinal, New England BioLabs) as per the manufacturer's protocol to prevent re-ligation. Both digested/dephosphorylated plasmid and PCR product were then separated in a gel and DNA extracted using Wizard Plus SV Miniprep DNA Purification Kit (Promega), and ligated overnight at 4.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were PCR screened for the presence of WPRE. All positive colonies were confirmed by sequencing using EGFP-I-F (Table 3). A list of all plasmids generated in this study and their purposes is in Table 4.

1.3.2 Transfection of Chicken Hepatoma Cells

[0103] The expression of EGFP was measured by transfecting CH-SAH cells with the dual-expression plasmids. Cells were seeded in 35 mm dishes at a density of 1.2.times.10.sup.6 cells/dish and incubated at 37.degree. C. with 5% CO.sub.2. Lipofectamine 2000 (Invitrogen) was used to transfect all constructs according to the manufacturer's recommendation. Briefly, 2 .mu.g of dual-expression plasmid and 5 .mu.l of Lipofectamine were incubated in separate 50 .mu.l aliquots of Opti-Mem medium (Gibco) for 5 minutes, then mixed and incubated together for 20 minutes. During this period of time, the medium was removed from each dish, and the cell monolayers were washed two times with phosphate buffered saline (PBS). Two ml of DMEM-F12 (5% FBS) without antibiotics was added to each dish. After 20 minutes, the plasmid-lipofectamine mixtures were added to the 35 mm dishes and incubated for 6 hrs at 37.degree. C. in the presence of 5% CO.sub.2. This was repeated for all ten dual-expression plasmids. After 6 hrs the medium was removed and fresh DMEM-F12 (5% FBS) was added. Every 12 hours post-transfection (h.p.t.), EGFP expression was confirmed by fluorescence microscopy and whole cell lysate was collected. Monolayers were washed with PBS, trypsin was added, and the cells were resuspended in DMEM-F12 (10% FBS). Cells were then centrifuged in 15 ml conical tubes (Nunc.RTM.), the supernatant was removed, and the cell pellet was resuspended in 500 .mu.l PBS and frozen at -80.degree. C. Transfected cell samples frozen at -80.degree. C. were freeze-thawed three times. The cell debris was spun down at 12,000 rpm in a microcentrifuge for 10 minutes at 4.degree. C. Supernatant was transferred to a fresh microcentrifuge tube and the protein concentration was determined at 280 nm using a Nanodrop 2000 (Thermo Scientific). All samples were adjusted to a protein concentration of 1 .mu.g/.mu.L.

1.3.3 Fluorescence Microscopy

[0104] Both transfected and infected CH-SAH cells were monitored by fluorescence microscopy with a Zeiss fluorescence microscope (Carl Zeiss) with FITC optics.

1.3.4 Measuring Reporter Gene Expression

[0105] The expression of EGFP was quantified by spectrofluorometry using a GloMax.RTM.-Multi (Promega) microplate reader. Briefly, 50 .mu.g of protein lysate was added in triplicate to a flat-bottomed black 96-well plate (Corning). Fluorescence of EGFP was measured in a GloMax.RTM.-Multi (Promega) microplate reader at 480 nm excitation and 528 nm emission wavelengths. The three readings were averaged to give one fluorescence value per sample.

[0106] Luciferase expression was determined using a Pierce Firefly Luciferase Glow Assay kit (Thermo Fisher Scientific) as per the manufacturer's protocol. Briefly, 25 .mu.g of protein lysate was added in triplicate to a flat-bottomed black 96-well plate (Corning). Fifty .mu.l of Luciferase assay substrate was manually added to each replicate, mixed well, and incubated for 15 minutes protected from light before measuring luminescence in a GloMax.RTM.-Multi (Promega) microplate reader. The readings were averaged to give one luminescence value per sample.

1.3.5 Normalizing Promoter Expression

[0107] Normalization of dual-expression constructs was performed to remove sample-to-sample variability of transfection efficiency (Schagat, T., Paguio, A., Kopish, K. 2007. Normalizing Genetic Reporter Assays: Approaches and Considerations for Increasing Consistency and Statistical Significance. Cell Notes. 17, 9-12). The fold change in activity was determined between promoter constructs and pCMV-EGFP-Luc. To begin, for each specific time-point and repetition the average fluorescence and luminescence value was determined for each plasmid construct, where a ratio of fluorescence over luminescence (F/R) was calculated. Activity level is being compared to the CMV promoter, currently used in recFAdVs, therefore the F/R ratio of CMV was set to a value of 1.0 (was divided by its own F/R ratio). Each remaining construct F/R value was also divided by the F/R value of CMV, resulting in a value representing the normalized fold change in activity (.DELTA.fold). Average fold change between repetitions was compared between all constructs at each time-point.

1.4 Generation and Characterization of Recombinant Viruses

1.4.1 Construction of Intermediate Constructs

[0108] Further analysis of EGFP expression in vitro was performed with recFAdVs containing the CMV, CAG, and Ef1.alpha. based expression cassettes. In previous studies, recFAdVs were recovered by homologous recombination between pFAdV-9.DELTA.4 and the PCR amplified expression cassette containing viral flanking regions, isolated from an intermediate construct. The plasmid pleft.DELTA.491-2,782 (pL.DELTA.2.4) contains the left end .DELTA.4 deletion site (.DELTA.491-2,782 nt) of FAdV-9 with a SwaI RE site for blunt-cloning a transgene (Corredor and Nagy, 2010b). In this study, a new intermediate construct system was developed by cloning viral flanking regions directly into the dual-expression plasmids, thus creating plasmids ready for recombination (pHMR). Viral genomic regions flanking the .DELTA.4 deletion site of FAdV-9 were PCR amplified from the intermediate construct pL.DELTA.2.4. The region left of the deletion site (VF1) was PCR amplified using 100 ng of pL.DELTA.2.4 and primers VF1-F and VF1-R (Table 3) with an annealing temperature of 52.degree. C. The resulting PCR product was gel extracted using the EZ-10 Spin Column Plasmid DNA Mini-prep kit (Bio Basic). Both VF1 PCR product and all dual expression vectors were digested with SpeI for 1 hr, followed by digestion with BglII. Both digested plasmid and PCR product were then separated in a gel and extracted using QIAEX II Gel Extraction kit (Qiagen), and ligated overnight at 16.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were PCR screened for the presence of the VF1 fragment with primers VF1-F and VF1-R. All positive colonies were confirmed by sequencing. Next, the region right of the deletion site (VF2) was PCR amplified using 100 ng of pL.DELTA.2.4 and primers VF2-F and VF2-R (Table 3) with an annealing temperature of 52.degree. C. The resulting PCR product was gel extracted with the EZ-10 Spin Column Plasmid DNA Mini-prep kit (Bio Basic). Both VF2 PCR product and all dual expression vectors positive for the VF1 fragment were digested with KpnI for 1 hr, followed by digestion with MfeI. Both digested plasmid and PCR product were then separated in a gel and extracted using QIAEX II Gel Extraction kit (Qiagen), and ligated overnight at 16.degree. C. Following transformation into E. coli DH5.alpha. cells and growth on LB-amp plates, colonies were PCR screened for the presence of the VF2 fragment using VF1-F and VF2-R primers (Table 3) at an annealing temperature of 52.degree. C., with an expected size of 2 kb plus the size of each EGFP expression cassette. A list of all six pHMR intermediate plasmids is in Table 4.

1.4.2 Generating Recombinant Fowl Adenoviruses

[0109] Recombinant FAdVs were generated to include the CMV, CMV-WPRE, CAG, CAG-WPRE, EF1.alpha., and EF1.alpha.-WPRE expression cassettes using a method modified from Corredor and Nagy (2010b) (FIG. 12). Expression cassettes flanked by viral DNA in the pHMR plasmids were recombined with pFAdV-9.DELTA.4 to create new recombinant FAdmids, containing a promoter of interest and EGFP. To obtain promoter EGFP cassettes flanked by viral DNA sequences, intermediate pHMR constructs were subjected to PCR or RE digestion with BglII/BamHI. Cassettes containing the CMV promoter (pHMR-CMV-EGFP and pHMR-CMV-EGFP-WPRE) and EF1.alpha. promoter (pHMR-EF1.alpha.-EGFP and pHMR-EF1.alpha.-EGFP-WPRE) were PCR amplified. PCR was carried out with 200 ng of plasmid and primers E1-F and E1-R (Table 3) with an annealing temperature of 52.degree. C., and the resulting PCR product was gel extracted using QIAEX II Gel Extraction kit (Qiagen). Cassettes containing the CAG promoter (pHMR-CAG-EGFP and pHMR-CAG-EGFP-WPRE) were double-digested using BglII/BamHI. Briefly, 5 .mu.g of plasmid was digested with both enzymes for 1 hr at 37.degree. C. Samples were separated by gel electrophoresis, and bands corresponding to CAG cassette (4.5 kb) or CAG-WPRE cassette (5 kb) were gel extracted using EZ-10 Spin Column DNA Gel Extraction kit (Bio Basic). Next, 5 .mu.g of pFAdV-9.DELTA.4 was linearized with SwaI and ethanol precipitated. The concentration of all DNA obtained by PCR and RE digestion product was measured with a Nanodrop 2000 (Thermo Scientific). For each construct, two .mu.g of promoter EGFP cassette and 2 .mu.g of SwaI digested pFAdV-9.DELTA.4 were co-transformed into E. coli BJ5183 cells. All resulting constructs were screened for recombination by NotI digestion followed by transformation into E. coli DH5.alpha.. These constructs were grown up in LB medium, isolated using PureLink HiPure Plasmid Midiprep kit (Invitrogen), digested with PacI, and extracted by ethanol precipitation. CH-SAH cells were seeded into 25 cm flasks at a density of 4.3.times.10.sup.6 cells/flask. Five .mu.g of PacI digested DNA was transfected into the CH-SAH cells using 25 .mu.l of Lipofectamine 2000 (Invitrogen). After 6 hrs the transfection mixture was removed and fresh DMEM-F12 (5% FBS) was added, the cells were incubated at 37.degree. C. with 5% CO.sub.2. Cells were monitored for the appearance of cytopathic effect (CPE) for the next week. At this point cell cultures were frozen and thawed three times, centrifuged, and the clarified supernatant containing recombinant FAdV-9.DELTA.4 was stored as P0 stock at -80.degree. C. Virus was subsequently passaged three times to obtain a high titre P4 stock. A list of recFAdVs is in Table 4.

1.4.3 Viral Growth Curves

[0110] One-step growth curves for all viruses were obtained as described by Alexander et al. (1998). Briefly, a total of 1.8.times.10.sup.6 CH-SAH cells were seeded in 35 mm dishes and incubated at 37.degree. C. with 5% CO.sub.2. The cells were infected with recFAdVs at a multiplicity of infection (MOI) of 5. After adsorption for 1 hour at room temperature, the cells were washed three times with PBS, and fresh DMEM-F12 (5% FBS) was added. Cell culture medium and cells were harvested at 0, 12, 18, 24, 30, 36, 48, 60, and 72 hours post-infection (h.p.i.). The medium was removed and frozen at -80.degree. C. as the extracellular virus, while the cells were washed three times with PBS, 1 ml of medium was placed on the monolayer and the dish frozen at -80.degree. C. as the intracellular virus. Extracellular virus was titrated for each time-point. CH-SAH cells were plated in 6-well plates at a density of 1.8.times.10.sup.6 cells/well and incubated overnight. Extracellular virus from each time-point was serially diluted (10.sup.-1-10.sup.-7), and 100 .mu.l of each aliquot was inoculated in duplicate and allowed to adsorb for 1 hr at room temperature. The inoculum was removed and the monolayer was washed in PBS. Three ml of agar layer consisting of 0.6% SeaKem LE agarose (Lonza), DMEM-F12 (5% FBS, L-glutamine, and PenStrep) was added to each well, and the plates were incubated at 37.degree. C. with 5% CO.sub.2. After five days, 1.5 ml of neutral red (0.015%) was added to each well, and after 24 hrs the plaques were counted.

1.4.4 Detection of EGFP Expression

[0111] A total of 1.8.times.10.sup.6 CH-SAH cells were seeded in 35 mm dishes and incubated at 37.degree. C. with 5% CO.sub.2. The cells were infected with recFAdVs at an MOI of 5. Uninfected and FAdV-9.DELTA.4 infected cells were the negative controls, while cells transfected with 6 .mu.g of pEGFP-N1 were the positive control. Cells were collected at 0, 6, 12, 18, 24, 30, 36, and 48 h.p.i. and centrifuged at 5,000 rpm for 5 minutes. Each sample was resuspended in 500 .mu.l of PBS and split into two 250 .mu.l aliquots. The first aliquot was stored frozen at -80.degree. C. to later be measured by spectrofluorometry. The second aliquot was centrifuged again to wash away FBS. The supernatant was removed and the cell pellet was resuspended in 200 .mu.l of RIPA lysis buffer (50 mM Tris HCl pH 7.5, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 10 mM EDTA, and 1% sodium deoxycholate). Samples were incubated on ice for 20 minutes, and re-centrifuged at 12,000 rpm at 4.degree. C. for 20 minutes. The supernatant was collected and stored at -80.degree. C.

[0112] EGFP expression was measured by spectrofluorometry at each time-point. Infected cell samples frozen at -80.degree. C. were freeze-thawed three times. The cell debris was spun down at 12,000 rpm for 10 minutes at 4.degree. C. Supernatant was transferred to a fresh microcentrifuge tube and the protein concentration was determined at 280 nm using a Nanodrop 2000 (Thermo Scientific). All samples were adjusted to a protein concentration of 1 .mu.g/.mu.L. For each sample, 50 .mu.g of protein extract was added in triplicate to a flat-bottomed black 96-well plate (Corning) and analyzed using a GloMax.RTM.-Multi (Promega) microplate reader to detect EGFP fluorescence using 480 and 528 nm excitation and emission wavelengths, respectively.

1.4.5 Bradford Assay and SDS-PAGE

[0113] Protein concentration was determined with the BioRad Protein Assay kit as per the manufacturer's protocol. Briefly, a 1:5 dilution of concentrated dye reagent was made in distilled H.sub.2O. Ten .mu.L of BSA protein standards, ranging in concentration from 100 .mu.g/ml to 1 mg/ml, along with whole cell lysate samples (diluted 1:10) were pipetted into a 96-well plate in triplicate. Two hundred .mu.L of the diluted dye reagent was added to each sample and mixed. After a 5 minute incubation, absorbance was measured at 595 nm in a microplate reader (BioTek Powerwave XS2). A standard curve was generated using the absorbance values of the BSA standards, and the equation of the trend line was used to extrapolate the concentrations of the unknown cell lysates. Values for each sample were averaged to generate an average total protein concentration, and dilution factor were accounted for. All samples were diluted to a final concentration of 0.67 .mu.g/.mu.L in distilled H.sub.2O.

[0114] Proteins were separated via SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Ten percent acrylamide gels were prepared according to recipes obtained from the Roche Lab FAQs Handbook (4.sup.th Ed). For all experiments, 15 .mu.L of each cell lysate (0.67 .mu.g/.mu.L) was mixed with 4 .mu.L of 4.times. SDS-PAGE loading buffer (supplemented with 2-mercaptoethanol). Samples were incubated at 95.degree. C. for 10 minutes prior to being loaded in the gels. A total of 10 .mu.g was loaded per sample into individual lanes, as well as 5 .mu.L of protein ladder (Precision Plus Protein Dual Colour, BioRad). Once all samples were loaded, gels were run at 100 V for approximately 1.5 hours in running buffer (25 mM Tris, 190 mM glycine, 0.1% SDS, pH 8.3).

1.4.6 Western Immunoblot

[0115] Following SDS-PAGE, proteins were transferred onto polyvinylidene difluoride (PVDF) membranes in a Mini Trans-Blot Electrophoretic Transfer Cell (BioRad) as per the manufacturer's protocol. Samples were run at 100 V for 1 hour in transfer buffer (25 mM Tris, 190 mM glycine, 20% methanol, pH 8.3). After transfer, membranes were rinsed in Tris-buffered saline supplemented with 0.1% Tween 20 (TBS-T) and blocked with 5% skim milk (in TBS-T) for 1 hour at room temperature with agitation. Primary antibody was added to the blocking solution and membranes were incubated overnight at 4.degree. C. To probe for EGFP, primary monoclonal mouse anti-GFP antibody (Molecular Probes) was used at a dilution of 1:1,000. To probe for actin, primary polyclonal goat anti-actin antibody (Santa Cruz Biotechnology) was used at a dilution of 1:1,000. The following day, all blots were washed three times in TBS-T for 10 minutes each with agitation. The blots were then incubated in secondary antibody diluted in blocking solution for 1 hour with agitation. The following polyclonal secondary antibodies conjugated with horseradish peroxidase (HRP) were used: goat anti-mouse (Invitrogen, at 1:5,000 dilution), and donkey anti-goat (Jackson, at 1:20,000 dilution). Blots were then washed three more times in TBS-T, and incubated in Western Lightning Plus-ECL reagent (Perkin Elmer Inc) for five minutes before being developed using a ChemiDoc XRS (BioRad).

TABLE-US-00002 TABLE 2 PCR conditions using either Kod or Taq polymerase as per the manufacture's protocol. Target size Step <500 bp 500-1000 bp 1000-3000 bp >3000 bp Kod Polymerase Activation 95.degree. C. for 2 min Denaturation 95.degree. C. for 20 sec Annealing Lowest Primer Tm.degree. C. for 10 sec Extension 70.degree. C. for 70.degree. C. for 70.degree. C. for 70.degree. C. for 10 s/kb 15 s/kb 20 s/kb 25 s/kb Repeat Steps 2-4 25 cycles Taq Polymerase Activation 95.degree. C. for 5 min Denaturation 95.degree. C. for 1 min Annealing Lowest Primer Tm.degree. C. for 30 sec Extension 70.degree. C. for 1 min/kb Repeat Steps 2-4 25 cycles

TABLE-US-00003 TABLE 3 Primers designed for the generation of dual expression constructs and reFAdVs. Restriction enzymes incorporated into each primer are underlined. Restriction Primer Sequence (5' to 3') Tm Enzyme pCl-Neo-F GGCTCATGTCCAATATGACCGCCAT (SEQ ID NO: 2) 62.degree. C. -- EGFP-F AAAAAAGAATTCACCATGGTGAGCAAGGGCGAG (SEQ ID NO: 23) 58.degree. C. EcoRI EGFP-R AAAAAAGCGGCCGCTTACTTGTACAGCTCGTCCATGCC (SEQ ID NO: 24) 59.degree. C. NotI EGFP-I-F GAGAAGCGCGATCACATGGT (SEQ ID NO: 25) 58.degree. C. -- .beta.-actin-F AAAAAAACTAGTTGGTCGAGGTGAGCCCCACGTT (SEQ ID NO: 26) 65.degree. C. SpeI .beta.-actin-R AAAAAAGAATTCGCCCGCCGCGCGCTTCGCTT (SEQ ID NO: 27) 71.degree. C. EcoRI L2R-F AAAAAAACTAGTCATAGTAAATATGGGTAACTTCTTAATAGCCA (SEQ ID NO: 28) 55.degree. C. SpeI L2R-R AAAAAAGAATTCATGGTTTGTTATTGGCAAATTTAAATAATGTT (SEQ ID NO: 29) 55.degree. C. EcoRI Luc-F AAAAAACCTAGGTTGGCAATCCGGTACTGTTGGT (SEQ ID NO: 30) 59.degree. C. AvrII Luc-R AAAAAATTCGAAGCCGCCCCGACTCTAG (SEQ ID NO: 31) 58.degree. C. BstBI SV40-F GGCCTCTGAGCTATTCCAGA (SEQ ID NO: 32) 56.degree. C. -- WPRE-F AAAAAAGCGGCCGCTCAACCTCTGGATTACAAAATTTGTGAA (SEQ ID NO: 33) 56.degree. C. NotI WPRE-R AAAAAAGCGGCCGCGCCCAAAGGGAGATCCGAC (SEQ ID NO: 34) 58.degree. C. NotI VF1-F AAAAAAAGATCTTACATGAATGACGCTGCTG (SEQ ID NO: 35) 53.degree. C. BglII VF1-R AAAAAAACTAGTAAATACACCGAGAAATACCACG (SEQ ID NO: 36) 53.degree. C. SpeI VF2-F AAAAAACAATTGAAATAAAGACTCAGAACGCATTTTCC (SEQ ID NO: 37) 54.degree. C. MfeI VF2-R AAAAAAGGTACCCCCCCGCAGAGAATTAAAA (SEQ ID NO: 38) 53.degree. C. KpnI E1-F ACATGAATGACGCTGCTG (SEQ ID NO: 39) 53.degree. C. -- E1-R CCCCCGCAGAGAATTAAAA (SEQ ID NO: 40) 52.degree. C. --

TABLE-US-00004 TABLE 4 Vectors and viruses in this study and their key features. pHMR-CAG-EGFP-WPRE 9,484 Intermediate construct for homologous recombination with pFAdV-9.DELTA.4 pHMR-EF1.alpha.-EGFP 8,766 Intermediate construct for homologous recombination with pFAdV-9.DELTA.4 pHMR-EF1.alpha.-EGFP-WPRE 9,290 Intermediate construct for homologous recombination with pFAdV-9.DELTA.4 pFAdV-9.DELTA.4 44,971 FAdmid backbone .DELTA.491-2,782 pFAdV-9.DELTA.4-CMV-EGFP 46,826 FAdmid containing CMV-EGFP in .DELTA.4 site pFAdV-9.DELTA.4-CMV-EGFP- 47,350 FAdmid containing CMV-EGFP-WPRE in .DELTA.4 site WPRE pFAdV-9.DELTA.4-CAG-EGFP 47,583 FAdmid containing CAG-EGFP in .DELTA.4 site pFAdV-9.DELTA.4-CAG-EGFP- 48,107 FAdmid containing CAG-EGFP-WPRE in .DELTA.4 site WPRE pFAdV-9.DELTA.4-EF1.alpha.-EGFP 47,389 FAdmid containing EF1.alpha.-EGFP in .DELTA.4 site pFAdV-9.DELTA.4-EF1.alpha.-EGFP- 47,913 FAdmid containing EF1.alpha.-EGFP-WPRE in .DELTA.4 site WPRE FAdV-9.DELTA.4 42,772 FAdV-9 deletion virus (.DELTA.491-2,782) Viruses Name Size (bp) Description FAdV-9.DELTA.4-CMV-EGFP 44,627 Recombinant CMV-EGFP virus FAdV-9.DELTA.4-CMV-EGFP- 45,151 Recombinant CMV-EGFP-WPRE virus WPRE FAdV-9.DELTA.4-CAG-EGFP 45,384 Recombinant CAG-EGFP virus FAdV-9.DELTA.4-CAG-EGFP- 45,908 Recombinant CAG-EGFP-WPRE virus WPRE FAdV-9.DELTA.4-EF1.alpha.-EGFP 45,190 Recombinant EF1.alpha.-EGFP virus FAdV-9.DELTA.4-EF1.alpha.-EGFP- 45,714 Recombinant EF1.alpha.-EGFP-WPRE virus WPRE

EXAMPLE 2

Generation and Characterization of FAdV-9 Viral Vectors

[0116] As shown in FIGS. 1-8, the inventors performed experiments to generate and characterize FAdV-9 viral vectors including ORF0-ORF1-ORF2 deleted viruses with an inserted mCherry coding sequence and using the native early promoter; TR2-ORF17-ORF11 deleted viruses with an inserted EGFP cassette and a dual expression virus useful as a polyvalent vector.

[0117] At the left end of the genome: from nucleotide 847 to 2753; 1906 nucleotides were deleted. This deletion includes ORF1A, ORF1B, ORF1C, and ORF2. At the right end of the genome: from nucleotide 38,807 to 42,398; 3591 nucleotides were deleted. This deletion includes TR-2, ORF 17 and ORF 11. The total deletion is 5497 bp; the size of the dual deletion vector is 39,567 bp. The foreign gene inserted to left end was mCherry (SEQ ID NO: 3, 711 bp). The foreign gene inserted to right end was EGFP (SEQ ID NO: 2, 1602 bp). Based on the 105% adenovirus stability rule, the capacity of dual vector is up to 7751 bp, this could be in different configurations, e.g. at the left end is up to 4160 bp and at the right end is up to 5844 bp.

[0118] As shown in FIGS. 2 and 3A, at the left end of the genome: from nucleotide 575-2753: 2178 pb nucleotides were deleted. This deletion includes ORF0, ORF1A, ORF1B, ORF1C and ORF2 and the foreign gene inserted to the left end was mCherry (SEQ ID NO: 3, 711 pb). The verification of FAdmid and the corresponding viruses were performed (FIGS. 3B and 3D) which show that the construction was correct. On the other hand, a CPE of the recombinant viruses was observed on CH-SAH cells, however the infected cells did not show any mCherry fluorescence which indicates that the foreign gene, in this case, mCherry, is not being expressed (FIG. 3C).

[0119] As shown in FIGS. 2 and 4A, at the left end of the genome: from nucleotide 847 to 2753; 1906 nucleotides were deleted. This deletion includes ORF1A, ORF1B, ORF1C, and ORF2. The foreign gene inserted to the left end was mCherry (SEQ ID NO: 3, 711 pb). The verification of FAdmid and the corresponding viruses were performed (FIGS. 4B and 4D) which show that the construction was correct. CH-SAH cells infected with the recombinant viruses showed mCherry fluorescence which indicates that the foreign gene, in this case, mCherry, is being expressed (FIG. 4C).

[0120] As shown in FIGS. 5 and 7A, at the right end of the genome: from nucleotide 38,807 to 42,398; 3591 nucleotides were deleted. This deletion includes TR-2, ORF 17 and ORF 11. The foreign gene inserted to left end was EGFP (SEQ ID NO: 2 1602 bp). The verification of FAdmid and the corresponding viruses were performed (FIGS. 7B and 7D) which show that the construction was correct. CH-SAH cells infected with the recombinant viruses showed EGFP fluorescence which indicates that the foreign gene, in this case, EGFP, is being expressed (FIG. 7C).

[0121] As shown in FIG. 8A, at the left end of the genome: from nucleotide 847 to 2753; 1906 nucleotides were deleted. This deletion includes ORF1A, ORF1B, ORF1C, and ORF2. At the right end of the genome: from nucleotide 38,807 to 42,398; 3591 nucleotides were deleted. This deletion includes TR-2, ORF 17 and ORF 11. The foreign gene inserted to left end was mCherry (SEQ ID NO: 3, 711 bp). The foreign gene inserted to right end was EGFP (SEQ ID NO: 2, 1602 bp). The verification of FAdmid and the corresponding viruses were performed (FIGS. 8B and 8D) which show that the construction was correct. CH-SAH cells infected with the recombinant viruses showed mCherry and EGFP fluorescence which indicates that the foreign genes are expressed (FIG. 8C).

[0122] FIG. 6 shows an additional vector based on the deletion of ORF17 located at the right end of the genome of FAdV-9 as part of the logical and systematic development of the vector system.

[0123] From the above, it can be observed that when ORF0 is deleted the native early promoter is not functioning since no foreign gene expression is observed (e.g. m-Cherry). However, the expression of genes reporter does occur when exogenous (foreign) promotor (CMV) is used (Corredor and Nagy, 2010b). When only ORF1 and ORF2 are deleted the native promoter works and the foreign gene reporter expression is observed.

[0124] FIG. 9 shows FadV-9 based recombinant viruses with reporter genes. Specifically, this figure is a summary and linear representation of FIGS. 4, 6, 7 and 8, wherein line pFAdV-9-RED represents FIG. 4, line pFAdV-9-.DELTA.17 represents FIG. 6, line pFAdV-9-EGFP represents FIG. 7, line pFAdV-9-Dual represents FIG. 8.

[0125] Accordingly, the inventors have successfully replaced ORFs 0-2, ORFs 1-2, and TR2-ORFs 17-11 of FAdV-9 with reporter genes. It was observed that FAdV left end early native promoter can drive foreign transgene expression. Furthermore, a large right end deletion in TR2 was demonstrated to be stable, contrary to previous studies. FAdV-9 with deletions at the left end and right end of FAdV-9 therefore may be used as polyvalent recombinant FAdV-9 viruses and may have applications for creating dual expression viruses suitable for vaccine vectors.

[0126] The primers used in this study are disclosed in Table 5.

TABLE-US-00005 TABLE 5 Name Sequence (5' to 3') Location Purpose ORF1-2Ver-F gcacagtcccaatggctt Pei et al., 2015 (SEQ ID NO: 42) ORF1-2Ver-R aaattctggtccgttaccga Pei et al., 2015 (SEQ ID NO: 43) TR2-17-11VerF GCCTACTCCTCAGCCTATCAC 38163-38184 verification (SEQ ID NO: 44) TR2-17-11VerR CAGTCCTCTAACGAGCACGC 43113-43133 verification (SEQ ID NO: 45) CAT-F GTGTAGGCTGGAGCTGCTTC Verification (SEQ ID NO: 46) ORF1CATSwaI-F ggacgtgctttaggtaatttcctccg 796-846 Deleted ORF1, (SEQ ID NO: 47) 1A, 1B, 1C, RF20 Ttctttttcccgtttaggtgaggag introduce SwaI (SEQ ID NO: 48) ATTTAATgtgtaggctggagctgcttc (SEQ ID NO: 49) ORF2CATSwaI-R gcgttctgagtctttattggtaa- Pei et al., 2015 (SEQ ID NO: 50) actcgaaacacgcgtcacacgcgc- (SEQ ID NO: 51) ATTTAAATcatatgaatatcctccttagttc (SEQ ID NO: 52) TR2-17-11CAT-F ccccctggcggccgttggccgccactg 38757-38806 Deleted TR2, (SEQ ID NO: 53) ORF17, ORF11 Cacccgcgccggacttagtctct introduce SwaI (SEQ ID NO: 54) TTTAAATgtgtaggctggagctgcttc (SEQ ID NO: 55) TR2-17-11CAT-R agtagagattataggaagcggggatta 42399-42448 (SEQ ID NO: 56) Tagtggtttttatttaaacgaga (SEQ ID NO; 57) ATTTAAATcatatgaatatcctccttagttc (SEQ ID NO: 58) ORF17CATSwaI-F tccccctggcggccgagggccgcca 40511-40561 Delete ORF17 (SEQ ID NO: 59) Ctgcacccgcgccggacttagtctct (SEQ ID NO: 60) ATTTAAATgtgtaggctggagctgcttc (SEQ ID NO: 61) ORF17CATSwaI-R gatacgaagaaggataggagcagaat 41461-41502 (SEQ ID NO: 62) Cgaggatacggtagttgactcc (SEQ ID NO: 63) ATATTTAAATcatatgaatatcctccttagttc (SEQ ID NO: 64) EGFPca-SwaI-F agctgcATTTAAATgtattaccgccatgcattag Pei et al., 2015 (SEQ ID NO: 65) EGFPca-SawI-R agctgcATTTAAATccacaactagaatgcagtg Pei et al., 2015 (SEQ ID NO: 66) mCherry-F agctgcATTTAAATATGGTGAGCAAGGGCGAGGAGG ??? amplify mCherry (SEQ ID NO: 67) mCherry-R agctgcATTTAAATCTACTTGTACAGCTCGTCCATGCCG ??? coding sequence (SEQ ID NO: 68) SwaI site are capitalized Sequences in bold are chloramphenicol cassette specific Underlined sequences are pN1-EGFP specific, the location is based on pEGFP-N1 Italicized sequences are extra nucleotides for SwaI digestion

EXAMPLE 3

Results

3.1 Expression of EGFP in Transfected CH-SAH Cells

3.1.1 Cloning Dual-Expression Constructs

[0127] A dual-expression system was created using EGFP and firefly luciferase (expressed under the SV40 promoter) to compare the strength of different promoter and enhancer elements on EGFP expression in vitro. The commercial plasmid pCI-Neo (Promega), containing the CMV promoter, was the backbone for the dual-expression system. The CMV promoter (944 bp) was subsequently removed using RE digestion with SpeI and EcoRI. After gel purification, both CAG (1,701 bp) and EF1.alpha. (1,507 bp) promoters were directionally sub-cloned into the pCI-Neo backbone using SpeI and EcoRI. Ligated product was transformed into competent bacterial cells and colonies were selected and screened by RE digestion (results not shown) and confirmed by sequencing. This process was repeated with both the .beta.-actin (285 bp) and L2R promoters (120 bp). The RE sites for SpeI and EcoRI were inserted into primers and both promoters were PCR amplified from plasmid DNA.

[0128] A 730 bp band corresponding to EGFP was PCR amplified with primers containing EcoRI and NotI sites. PCR product was directionally cloned into each promoter plasmid, transformed into competent bacterial cells and confirmed by both PCR (data not shown) and sequencing. Finally, firefly luciferase (1,712 bp) was PCR amplified with primers containing the RE sites AvrII and BstBI. Plasmid DNA, containing EGFP under the control of each promoter, was then digested with AvrII and BstBI to remove the neomycin resistance gene, and luciferase was directionally cloned in its place. The presence of luciferase in each plasmid was confirmed by PCR (results not shown) and sequencing. This resulted in the dual-expression plasmids: pCMV-EGFP-Luc, pCAG-EGFP-Luc, pEF1.alpha.-EGFP-Luc, p.beta.actin-EGFP-Luc, and pL2R-EGFP-Luc (FIG. 11). Five additional plasmids were generated to include the enhancer element WPRE. A 543 bp band corresponding to WPRE was PCR amplified with primers containing a NotI site. Each dual-expression plasmid was linearized with NotI and the WPRE element was non-directionally cloned into each. Proper orientation was screened by PCR (data not shown) and plasmids were confirmed by sequencing, resulting in the dual-expression plasmids: pCMV-EGFP-WPRE-Luc, pCAG-EGFP-WPRE-Luc, pEF1.alpha.-EGFP-WPRE-Luc, p.beta.actin-EGFP-WPRE-Luc, and pL2R-EGFP-WPRE-Luc (FIG. 13).

3.1.2 Expression Patterns of Promoters Observed by Fluorescence Microscopy

[0129] CH-SAH cells were transfected with the dual-expression constructs to follow EGFP expression patterns over 72 h.p.t. by fluorescence microscopy (FIG. 14). The earliest expression of EGFP was noted at 12 h.p.t. by CMV, CMV-WPRE, CAG, and CAG-WPRE plasmids, with expression levels increasing over-time. Both EF1.alpha. and EF1.alpha.-WPRE constructs began expressing EGFP between 24 and 36 h.p.t. and remained steady in expression over-time. Constructs containing the .beta.-actin or L2R promoters expressed EGFP the weakest in CH-SAH cells, with expression starting at 36 and 60 h.p.t., respectively. All constructs were compared to mock transfected cells (negative control) and a positive control of pEGFP-N1 transfected cells (data not shown).

3.1.3 Normalized Expression of Promoter Constructs

[0130] Dual-expression constructs were transfected into CH-SAH cells and transgene expression was measured over 72 h.p.t. The fluorescence of EGFP was measured by fluorometry at each time-point, while the luminescence from luciferase was measured using a Peirce Firefly Luciferase Glow Assay kit (Thermo Fisher Scientific). To better analyse the expression of EGFP driven by each promoter/enhancer element, and to minimize sample-to-sample variation, the expression of luciferase from each sample was used to normalize the results (Schagat et al., 2007). The data was analysed by calculating the fold change of each construct, at each specific time post-transfection, in relation to the CMV promoter (Table 6). The normalized expression of each construct from 12-72 h.p.t. is shown in FIG. 15. At 12 h.p.t. significant decrease in expression was observed by EF1.alpha., EF1.alpha.-WPRE, .beta.-actin-WPRE, and L2R-WPRE constructs compared to CMV. CMV-WPRE and CAG constructs had increased expression at 12 h.p.t. however this was not significant. Greater differences in expression patterns were observed between 24-72 h.p.t. In general, all constructs showed decreased activity over-time compared to the CMV control. At both 12 and 24 h.p.t. the CMV-WPRE construct showed an increased fold change of 1.047 over CMV only, though the differences were not significant. Between 36 and 60 h.p.t. the CMV-WPRE showed a slightly decreased fold change ranging from 0.851 to 0.922. A significant decrease in expression was deleted at 72 h.p.t., when CMV-WPRE showed a lower activity level of 64% compared to the CMV control, and this difference was significant. For each of the remaining constructs, expression was significantly below that of CMV at all time-points where P<0.05. Constructs containing the CAG and CAG-WPRE cassettes consistently expressed at a range of approximately 50% and 40%, respectively, compared to CMV from 24 h.p.t. Similarly, constructs containing the EF1.alpha. and EF1.alpha.-WPRE cassettes were found to have a decreased activity level compared to CMV, expressing at a range of approximately 20%. The remaining constructs, containing the .beta.-actin and L2R promoters, had activity levels less than 10% that of CMV.

3.2 Generation of Recombinant FAdV-9.DELTA.4s

[0131] 3.2.1 Cloning pHMR Constructs

[0132] To further analyse promoter activity and the activity of WPRE in the context of FAdV replication, recombinant FAdVs containing the most efficient EGFP expression cassettes were generated following a method modified from Corredor and Nagy (2010b). A 477 bp fragment (VF1) left of the FAdV-9.DELTA.4 deletion site was PCR amplified and directionally cloned into pCMV-EGFP-Luc, pCMV-EGFP-WPRE-Luc, pCAG-EGFP-Luc, pCAG-EGFP-WPRE-Luc, pEF1.alpha.-EGFP-Luc, and pEF1.alpha.-EGFP-WPRE-Luc. Subsequently, a 1609 bp fragment (VF2) right of the FAdV-9.DELTA.4 deletion site was PCR amplified and cloned into each plasmid, resulting in the intermediate plasmids: pHMR-CMV-EGFP, pHMR-CMV-EGFP-WPRE, pHMR-CAG-EGFP, pHMR-CAG-EGFP-WPRE, pHMR-EF1.alpha.-EGFP and pHMR-EF1.alpha.-EGFP-WPRE (FIG. 16). Successful cloning was determined by PCR screening and sequencing.

[0133] Intermediate constructs were subjected to PCR or RE digestion with BglII/BamHI to isolate promoter EGFP cassettes flanked by viral DNA fragments for use in homologous recombination with pFAdV-9.DELTA.4 (FAdmid). The resulting FAdmids were generated upon homologous recombination: pFAdV-9.DELTA.4-CMV-EGFP, pFAdV-9.DELTA.4-CMV-EGFP-WPRE, pFAdV-9.DELTA.4-CAG-EGFP, pFAdV-9.DELTA.4-CAG-EGFP-WPRE, pFAdV-9.DELTA.4-EF1.alpha.-EGFP, and pFAdV-9.DELTA.4-EF1.alpha.-EGFP-WPRE. Successful recombination was determined by sequencing and NotI digest screening (FIG. 17). Digested pFAdV-9.DELTA.4 resulted in a 4 kb, 5.1 kb, 13.7 kb, and 23.8 kb band. FAdmids containing the CMV cassettes were identified by a diagnostic 2.2 kb band corresponding to the CMV promoter plus the EGFP gene and SV40 polyA tail. FAdmids containing the CAG cassettes were identified by a diagnostic 2.9 kb band, and EF1.alpha. by a diagnostic 2.7 kb band. Additional bands at 550 bp were present in FAdmids containing the WPRE element. These FAdmids were transfected into CH-SAH cells and to generate the corresponding viruses.

3.2.2 Viral Growth Kinetics

[0134] Viral growth kinetics were compared among the six recombinant viruses and the reference FAdV-9.DELTA.4 to determine whether the addition of any promoter EGFP cassette affected virus replication and growth. The virus titer and growth of all recFAdVs appeared to follow a similar pattern to FAdV-9.DELTA.4, except that FAdV-9.DELTA.4-CAG-EGFP and FAdV-9.DELTA.4-EF1.alpha.-EGFP grew to a titer one half log lower than the reference starting at 24 h.p.i. (FIG. 18). Viruses grew to the following titers; FAdV-9.DELTA.4=2.4.times.10.sup.8 pfu/ml, FAdV-9.DELTA.4-CMV-EGFP=4.8.times.10.sup.8 pfu/ml, FAdV-9.DELTA.4-CAG-EGFP=8.3.times.10.sup.7 pfu/ml, FAdV-9.DELTA.4-EF1.alpha.-EGFP=8.2.times.10.sup.7 pfu/ml, FAdV-9.DELTA.4-CMV-EGFP-WPRE=3.1.times.10.sup.8 pfu/ml, FAdV-9.DELTA.4-CAG-EGFP-WPRE=3.0.times.10.sup.8 pfu/ml, and FAdV-9.DELTA.4-EF1.alpha.-EGFP=3.7.times.10.sup.8 pfu/ml. The appearance of CPE (FIG. 19) and plaque morphology (not shown) of recFAdVs were similar to FAdV-9.DELTA.4 in that the cells rounded and detached by 5-7 d.p.i. All recFAdVs expressed EGFP as seen by fluorescence microscopy.

3.3 Promoter Activity During Infection

3.3.1 Measuring EGFP by Spectrofluorometry

[0135] EGFP expression by recFAdVs was measured between 0-48 h.p.i. in CH-SAH cells. Based on both spectrofluorometry and fluorescence microscopy, expression of EGFP from all recombinant viruses was low until 12 h.p.i. Strong expression of EGFP was noted between 12-30 h.p.i. but it declined after 36-48 h.p.i. Maximum fluorescence readings were measured at 30 h.p.i. for all recFAdVs (FIG. 20). Cells infected with FAdV-9.DELTA.4-CAG-EGFP showed the greatest increase in EGFP expression compared to FAdV-9.DELTA.4-CMV-EGFP at all time-points, ranging from 12 (48 h.p.i.) to 29-fold (24 h.p.i) increase in expression. FAdV-9.DELTA.4-EF1.alpha.-EGFP infected cells also had increased expression compared to FAdV-9.DELTA.4-CMV-EGFP at all time-points, with a slightly lower range of 2 (12 h.p.i) to 20-fold (24 h.p.i) increase in expression. Viruses that contained a WPRE element expressed EGFP at a lower level when compared to their promoter counterparts. FAdV-9.DELTA.4-CMV-EGFP-WPRE was the "worst performing" virus, with 2.4 (48 h.p.i.) to 17-fold (36 h.p.i.) decrease in expression compared to FAdV-9.DELTA.4-CMV-EGFP. Although both FAdV-9.DELTA.4-CAG-EGFP-WPRE and FAdV-9.DELTA.4-EF1.alpha.-EGFP-WPRE performed worse than their promoter counterparts, both viruses on average expressed EGFP at a higher level than FAdV-9.DELTA.4-CMV-EGFP. FAdV-9.DELTA.4-CAG-EGFP-WPRE showed increased expression ranging from 1.3 (48 h.p.i) to 4.3-fold (24 h.p.i.), while FAdV-9.DELTA.4-EF1.alpha.-EGFP-WPRE expression ranged from a 2.5-fold decrease (12 h.p.i.) to 3.6-fold increase (24 h.p.i.).

3.3.2 Expression of EGFP Measured by Western Blotting

[0136] In addition to spectrofluorometry, viral EGFP expression was also examined by Western immunoblot. Whole cell lysate was collected from infected cells at 0, 6, 12, 18, 24, 30, 36, and 48 h.p.i. At each time-point, 10 .mu.g of total protein from each lysate was separated by SDS-PAGE, blotted, and the blot was probed using the anti-EGFP antibody (FIG. 21). The molecular mass of EGFP was expected to be 27 kDa (Takebe, N., Xu, L., MacKenzie, K. L., Bertino, J. R., Moore, M. A. S. 2002. Methotrexate selection of long-term culture initiating cells following transduction of CD34+ cells with a retrovirus containing a mutated human dihydrofolate reductase gene. Cancer Gene Ther. 9(3):308-320). At early time-points of 0 and 6 h.p.i., no EGFP signal was detected from any recFAdVs. By 12 h.p.i., a band corresponding to EGFP was detected in lysate collected from FAdV-9.DELTA.4-CAG-EGFP infected cells, which was detected up until 48 h.p.i. Cell lysates collected from FAdV-9.DELTA.4-EF1.alpha.-EGFP, FAdV-9.DELTA.4-CAG-EGFP-WPRE, and FAdV-9.DELTA.4-EF1.alpha.-EGFP-WPRE also produced a detectable EGFP band from 18-36 h.p.i. In contrast, no EGFP bands were seen for FAdV-9.DELTA.4-CMV-EGFP and FAdV-9.DELTA.4-CMV-EGFP-WPRE infected lysates at any time-points. The negative controls, FAdV-9.DELTA.4 infected lysate and mock infected lysate, showed no signal for EGFP throughout the time-course. Transfected CH-SAH cells containing the EGFP positive plasmid under the CMV promoter, pEGFP-N1, provided a positive signal from 12 h.p.t. onwards. At each time-point blots were probed with an anti-actin antibody to show relative levels of equal loading of all samples. Actin levels appeared similar at each time-point, suggesting equal loading, however at 48 h.p.i. actin was not detected in virus infected lysates.

TABLE-US-00006 TABLE 6 Fold change in promoter activity of transfected CH-SAH cells at various hours post-transfection in relation to the CMV promoter. Significant activity is determined by a P value <0.05. Time-point (h.p.t.) Promoter .DELTA.fold SD P value 12 CMV 1.0 -- -- CMV-WPRE 1.047 0.245 0.7526 CAG 1.483 1.735 0.5506 CAG-WPRE 0.479 0.370 0.0717 EF1.alpha. 0.610 0.184 0.0015 EF1.alpha.-WPRE 0.313 0.117 0.0005 .beta.-actin 0.772 0.418 0.2587 .beta.-actin-WPRE 0.357 0.097 0.0003 L2R 0.566 0.528 0.1654 L2R-WPRE 0.134 0.095 0.0001 24 CMV 1.0 -- -- CMV-WPRE 1.047 0.116 0.5177 CAG 0.409 0.116 0.0001 CAG-WPRE 0.294 0.039 0.0001 EF1.alpha. 0.156 0.076 0.0001 EF1.alpha.-WPRE 0.087 0.018 0.0001 .beta.-actin 0.074 0.042 0.0001 .beta.-actin-WPRE 0.042 0.043 0.0001 L2R 0.028 0.016 0.0001 L2R-WPRE 0.010 0.002 0.0001 36 CMV 1.0 -- -- CMV-WPRE 0.894 0.156 0.3078 CAG 0.560 0.331 0.0180 CAG-WPRE 0.387 0.139 0.0016 EF1.alpha. 0.196 0.057 0.0001 EF1.alpha.-WPRE 0.124 0.063 0.0001 .beta.-actin 0.056 0.037 0.0001 .beta.-actin-WPRE 0.015 0.007 0.0001 L2R 0.014 0.013 0.0001 L2R-WPRE 0.002 0.001 0.0001 48 CMV 1.0 -- -- CMV-WPRE 0.922 0.135 0.3758 CAG 0.431 0.036 0.0001 CAG-WPRE 0.303 0.050 0.0001 EF1.alpha. 0.187 0.063 0.0001 EF1.alpha.-WPRE 0.085 0.022 0.0001 .beta.-actin 0.032 0.017 0.0001 .beta.-actin-WPRE 0.013 0.005 0.0001 L2R 0.015 0.014 0.0001 L2R-WPRE 0.001 0.001 0.0001 60 CMV 1.0 -- -- CMV-WPRE 0.851 0.123 0.1039 CAG 0.572 0.257 0.1423 CAG-WPRE 0.392 0.191 0.0053 EF1.alpha. 0.254 0.054 0.0026 EF1.alpha.-WPRE 0.124 0.021 0.0001 .beta.-actin 0.033 0.007 0.0001 .beta.-actin-WPRE 0.015 0.004 0.0001 L2R 0.002 0.001 0.0001 L2R-WPRE 0.001 0.001 0.0001 72 CMV 1.0 -- -- CMV-WPRE 0.643 0.056 0.0004 CAG 0.591 0.137 0.0521 CAG-WPRE 0.434 0.164 0.0040 EF1.alpha. 0.296 0.082 0.0067 EF1.alpha.-WPRE 0.234 0.119 0.0004 .beta.-actin 0.030 0.002 0.0001 .beta.-actin-WPRE 0.028 0.018 0.0001 L2R 0.002 0.001 0.0001 L2R-WPRE 0.003 0.003 0.0001

EXAMPLE 4

Other Embodiments of Recombinant FAdV-9

[0137] FIG. 22 shows other embodiments of recombinant FAdV-9 wherein ORF1 and 2 are deleted at the left end of the genome and replaced with HA of H7 coding sequences and wherein ORF19, ORF11 or TR2 are deleted at the right end of the genome and replaced with a HN cassette, HN-IRES-HA of H5 cassette or HA of H5 cassette, wherein IRES (Internal Ribosomal Entry Site) allows for translation initiation in the middle of a messenger RNA (mRNA) sequence as part of the greater process of protein synthesis. The IRES used (SEQ ID NO: 41) is from a Canadian isolate of avian encephalomyelitis virus isolate (AEV-IRES)

[0138] While the present disclosure has been described with reference to what are presently considered to be the preferred examples, it is to be understood that the disclosure is not limited to the disclosed examples. To the contrary, the disclosure is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

[0139] All publications, sequences, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, sequence, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

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Sequence CWU 1 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 68 <210> SEQ ID NO 1 <211> LENGTH: 45063 <212> TYPE: DNA <213> ORGANISM: Fowl aviadenovirus D <220> FEATURE: <221> NAME/KEY: whole genome <222> LOCATION: (1)..(45063) <400> SEQUENCE: 1 catcatctat atatacctac atgaatgacg ctgctgtcac ctcgcactac cgctcccgcc 60 cgctagatgg caaaattgct ccaccaaagt atttccgtga tacaaagtcc ggactgggcg 120 cggcgaatac gtgccgggcg aggcacggag ccgactttct ggcaccgata gcaataaatg 180 ggcgttccct tgcgtgcgga attgcgccgg cacagtccca atggcttggg agcggtccag 240 aaagcgatta gtttttgcgt acgcacgcga aagctatttt tgcagcactt acttccttat 300 ttgtttacaa ccgcctccgc ctgtgtgtaa tttaaaattt gtacataccg ttccgatcgc 360 cgtacttggg cttggaacgc ggacttcgag tccctaaatt gatttatggt gtcttaatca 420 gcgctttttg tgccacgttg tgcgttctcc gttgaccttt gaaaaggcgc gtggtatttc 480 tcggtgtgtt tttcgggcgg attacgcgtt acgtgatcgt tttatggcgg gctccgggcg 540 tataacagtg gagcggctga tcgtcagagc acggatgagc gagcagccgg actcaccgcc 600 ggagctctat ccgtctctcc tggtcctgcg ggtgaacccg gttccccgta ctccgttcgt 660 tttggacatc agccgtttgc ccgtggtgcc ggagcgcatt ttgctgcacg actttccgaa 720 cggaccctgg tcggaaacga tttggtacgg gaagatcact cagcgggagc tgaaccaggc 780 gctagagtct atcgtggacg tgctttaggt aatttcctcc gttctttttc ccgtttaggt 840 gaggagatgt ctttcgattc cggttgcccc ccgactccac cggtaaagct gctgttcaag 900 aagcattccc cgtttgccgt gacgccccag cgtgctacct cgggggctgc cggttacgac 960 ttgtgcagct cggcggacgt ggtggtgccg ccgaagagcc ggtctctcat tccgaccgac 1020 ctttcttttc aattcccccg aggcgtgtac gggcgcatag cccctcgttc gggcttggcc 1080 gtaaagtttt ttatcgatgt gggagccggt gttatcgata gcgattaccg gggcatcgtt 1140 agcgtcctcc ttttcaattt ttcggaccac aatttcaacg tccgtcgggg cgatcgcata 1200 gctcagttaa ttttggagcg tcatttgacg cccgacttgg aagagcgatc gggcttagac 1260 gagacggcca ggggtgcagc cggtttcggc tccaccggcg gttttgatac aggggtctgt 1320 ccctcctcct tttcttagga attgaccgag agcgaggcgg cggacgaagt ggatttcgcc 1380 gttcaactgg cggccgctct cgaggcagat aacgaggcta ctgcgggttg cttctgcgaa 1440 gccgactccg ggaaaaacac ctgttctctg tgttcttgga actgtatcat tctttaggta 1500 tgattgtttc tgtttggacc ctcgttgtag agctcgttaa cgcgtgggat cgtttgtgtg 1560 gtcgtcgggc ttaccgctat tacgcggatc gcgagctgtt gtggcaagtg cacgagagtc 1620 tgcggtccgc cttggagaag acggtcggtg tcggaagagg cttggcccgc atcggcgacg 1680 gtgaaaacgg gtatttcgga ggaggcggcg aagctgttcg cgacaattga cgacatgtcg 1740 atcgaagatt ggttcctcgt tttcgtagtg cttttcgcta ttctgtcttt tatgtggggg 1800 atgttcttgg ggatgctttg cgggccaggc gatttggatt tcgcgggtta ctgggactgg 1860 tatttccgtg tttgtaacgc gagggttcct taataaatgt cttttcgtaa cgttaattcc 1920 gcgtttgggt tttgcttttt gcagacggga tgtcgggctc gagggatcgc tattggtcct 1980 tggtcgaatc tctgataaat agagggatcg tgacccgtca gcagtggtat ttcaacgacc 2040 gctccgagta cgggtattac acgaagcgct acagaggcgc gatgaaattg tccaacttgt 2100 tccgcgacgt gcgtcgtcac atgttctgga caaaatgttt ggccgactac atgttgaccc 2160 ccgaaaccga ctttcttcaa atcgaggaga atccgtttta ttcggtgttt tcgaaaaacg 2220 gctacgatcc cgctctcgtg ggctcgatgt tgctggcttg ggccgcgcgg cagtcgaagc 2280 gcaataccgt ttggattagg ggagcgcccg tgacgggagc tccttacttg gcggaagcga 2340 tcgcttactg ctgcccgttg atgacgtgcg tcgattggcg caacaagcag aatccttttg 2400 agaaggggtt cgagtctttg ctgttttggt gggacggcgg gcacgtgccg caccgctgcg 2460 tggccttgtg tcgccaggtg tttaggggcg aaaacgtcat gtttccccgg gaagacggtc 2520 aaatgagcga attggtgcgt acgcccgtgt tgttgtactc tgaacacgac gcttgtcgca 2580 cgttagtcaa gtggggcgtg ttcagtgatg agttttctga cggtttacgt aattccatgt 2640 attgtttaga cttgtccgcg cccatggaag gcgctcagtg cgtcacttgt gccgacgctc 2700 gcgcgttttt tacgtgggct tacccgcgac gcatggattt tgactgtttt tgagcgcgtg 2760 tgacgcgtgt ttcgagttta ccaataaaga ctcagaacgc attttccgtg tgaccgatgc 2820 tttttattgt taaatttcaa cgggaaagct tgcaaaacgg atagcatatg acgactagcc 2880 taaaggggaa ggtttcgagg ttgtcctgga cgcgacgaca acagcgcatt aagctgttgt 2940 taccgtagga gggagtgatg aatatctcgt aaccgcctcg tatgcgggat aaggcgtaat 3000 tattgcggac ggcgcggtgg gtcagcaagt cgtgcagagg ggccagaggg ttcaggatgt 3060 agtcgagtcg tcccgtatcg tcgtcggggg cggtggagat gagtaagagg ttgaaattgg 3120 tgatgttaac gagaggatag gggttgttcg cgcgctcttc ttcaaagtac tcgtacactt 3180 tatccaacca gtatgtgcac acgactttgc tcgctgcttg acacatgagg ctgtagggat 3240 tggtgcaggt gcaatcgtat cggatttgag gcaaggggga ttgctgctcg aggtcttcgg 3300 taacggacca gaatttattg tacagacgac agcgcgccct gcatttgggc atgctgtatt 3360 cgggcagtat ttcttcgaag gtgacggtgt ccttcctggg gaacggaaac atgcgttctc 3420 cgtgaagaaa ctctaagaaa aagatgtcaa gagcgaggtg agcgggtgct tccagctggg 3480 cttctttgca ttctagcgtg agagagtgca cgctgtcctg cggaaagaga gaaaattaca 3540 ggaacacggt gatgacgggg tacgggaact tttgcatgag tctcaggaag cacggtatga 3600 gtgtgtaatc cctgtgcgac cgcggtaacg cgatgcgaat gtgtccgtcg accacttcta 3660 tcttggcgaa gggtttggag gaatacacga ataggttgat gacgccgtcg atgatttcga 3720 tgggaggtct gccgtgacat cggaagcagt gcgtgcaacg ttttcggacg tattggtcga 3780 aatggatggg aaacgggttt tccgcgatgg ggtacgcggt gagtttggcg tgccactcca 3840 tggtgagtgt ggcggctagg gactgacaaa agagcgagag cggctcctcg cactcgcata 3900 cgtagctcgc gacccattct cccatccatt tccgacctcc gtcctctacg tcggctaata 3960 ggttgctaat tttaaaggag gactggaacg gcctcagcct gaacggcagg gattcggggg 4020 taaagtaggg ctctgaccga tagggataca tggatcttcc gcataggaaa cgcagtagat 4080 ccggatcgaa aagggccaag gccgacacga cccgcagggg cgaggggaat tttacggagt 4140 agacgtgttc gcaggtgtca ccgaggttgt cgctgtccca tcttatgtga aaagtggacg 4200 aataacgaga gggagacgag atctgcaaag aaaagaaagt tttattgggg gataactttt 4260 tcagtattcg ggagagtagc ccgcgcaagt ctgacaaagc ctgttttcgc gcttgcagtg 4320 catgaccgct gcgcaaggta cggttctgga ggtcgacgcg acaaagtctt ttaattctct 4380 gcgggggttg tggcagacgg gggtgagctc ttggggagcc gtgaggaaca ggatggtcga 4440 gtttctgcac aggaacgaac aggcgataga gacgtcggtg caatggatga gcatttgggg 4500 cgctctgacg aagacggggc gaccggcgat gtagcaggtg cccgaattgc cctccataaa 4560 gtgtagcgcg agcgtggacg gggctctgtc gacgaagggc acgcagtaga gcgacgtgtg 4620 ttgggcgacg ttggctatcc gctctatgtc gttgagatcg ctatctgaga tggccattgg 4680 gaagcaggag gcgaggacgt tgtacagcag cttggcgttg gcgaggcgtc ctccgcacag 4740 gatgagcgta ttgatctctc cgttgagcca ccgtttgatg gtgagggcgc acagagccgc 4800 gtcgtacccc tctttttcga gcagctgatg gaaccagttc tcttcgggat tgtagaaagc 4860 gagtctgagg gactcgggtt cctcgtaggg cagttgggcg tagagagatc tggagtttcc 4920 gaatctttct cggagggccc ctaggacgta ttcccgctcg tccggtctga gggtgtactg 4980 gtcgtaggcg ggcatctggt cccactgaga tcggtgagcg atgccgtctc ggaaaagccg 5040 ttcgatgagc gcgtccatct gcgacgccga cctgttggtc tcctgagagt agcgggaagg 5100 agtctttttg ttcggatcgg gtgtgggcgg agtttctggg acgtaaggaa agccggagaa 5160 gtcgtcttcg tcgtcctccg tgcgcctctt ctgcgacggg ggcggcgagg agggagaggg 5220 actacgatgt tccgctttcc gcttactctg gaagagagag gatatgatca aacttctgct 5280 tgcaggagta acacggggga tcgatccagg cggtggagca gagcactccg aaatcgtttt 5340 tgcagtcaaa atcgcgttcc ccttcgtgct cgcgtatctg cctcaggtat tgcacgagct 5400 cgctggtgag gaaggagatt ccgtcgtcgt tggcggtttc gaactggatg atgaggtggt 5460 actttgggtt ggtgggtagt tggtctgggc gcggcagctt gttgagtcgc acgacgcact 5520 tcacttcttc gatttcgacg agttcgccct gatgcgagac gttgaaagcc ctgcccctga 5580 gcatttcgtt gacgagtggg tttttaaagg ggtactcgtg atgcaggggc gggaagtaga 5640 tgagtttagt gacgtctcgc acgggaacgt actggtcgaa atcgaagttg ttgacgtgcg 5700 cggtcaggac gcagtggaac atgcgcaaga tagagttggt aaaggtgtct gcgcaggtgg 5760 aggagtcgcc cacaatgtag agaacgttga gagtgtcgtg caagcgcgaa agcgtgtagt 5820 cgctgagcca gcgaccgagc atgtagctta gggtgacgag attgaaccct tccctttccg 5880 cgaatcttgc gagatgcctg ctggtttcta tggtgaaggg gccgctaggg ctattgagta 5940 gggtttggta gaggctaaag ttgcagtaga ttttcctggc ttcgtggaga agcagttttc 6000 cctgcagacc tactccgtat cgagcgaatt gctccggatc cctgagacgc cattctcctt 6060 cggtgaaaat gagcctcttt tggagctggt aggcgagttc gcccggattt tcgggggcgt 6120 tcgaccgctt attggaaccc gtttgcctcg acctgcgact ctggagggga agacaggggt 6180 tcgaagacgg ttagtttagg gcgcttggcg agcatgtttt tgcgagcctc tgcgaatcgt 6240 atgatgagct tggaggctct gaagaatttt tcctgcacgt tgatgttaag gggcaggatg 6300 gcgtaccgct gatcgagcac gcaccagcgg aacgagtcgg agacggggtc gggggagtaa 6360 gtgagccagc tgaacttttg gttctgctgg agaaaagcga agtaggcttt gagtaacacg 6420 accagttctt tgctgatgtt gtgggcgtaa gagtagagaa agcgcgagaa ctgaaattgg 6480 gggatgtggc aagagaggat gtgtattttg gcgttgacct taagggtcgg cacgtttcct 6540 atggcggtgc gcggcgcgag attgtggaga accacgaaaa cgtagaaagc ggtacagtta 6600 gcggaacgag cgaagagctt agaggggaga gcgtgaaaga gcacggagac gctagacccg 6660 gagcagagtt tatccatgca ctcgtccatg acgatggcca gcggtccctg ccgggacgcg 6720 ttgacgtata cgttgtccgg gtgttcgatg ttgaggtttt cgggcgaggt tgcttcctcg 6780 tacgtcattt ctacgaactc cggtcggaag gtgcaggttc tgggcgcgaa ggttccgtcc 6840 tccttgcagt cgtagttgga ttccagaagt tgtaggttcc aagccgtttg ttccaccggc 6900 gggatcatat tcttctccgg cgtcacaaag ataaccgttt ccgggatcgg ttgcagcata 6960 tcgcaggaga tcaacgctct gagcagatgg ctcttgcccg atcccgtcgg gccgtagatg 7020 acgccgatga cgggctgtcg gcccatgttg agcgacggta gttgtccgtc gacgaggtac 7080 cgactgtctt cctgttcttg ccgttccacg gctagctgta tttctaggaa cttgtcgtcc 7140 actcccccga gactgtaaaa gtcgtcgaag ctgggaaatt tctgttcgcg aaagagatgt 7200 ggggctaggt ccgccgctcc ttcgtaccag cccgttaccc gtttgtagaa ctcgcctgcc 7260 cgtagatact cttcctctcg gtagcgccat gttttcctct ttttattagc gacggccccg 7320 gcctccgctc cgctcatggg tctccgagga gatccaaaat ttcgtcacag tcctcgatag 7380 agagcgaccg gtctgggacg tcgaactcgt ccgagtcctc tccgtcgtcc tcgggagcga 7440 tccgtgcggc cgccgccgcc gcgagcagcc tgaggggggc tagaggttct tcgtggccca 7500 ccgggggcac tctccggact tcgtcggcgg ttctcgggtt gggatgttcg gtgtcgtagg 7560 ggtaaagaaa gtccccgtgg cgatactgcg tgaggtcctt ccagggtctt aggactctgg 7620 tgagctgttc gttgtggatg gtgaacggat cgtatcggct caccttgttg aggagcgtgg 7680 ttttgaagat ggtcctgcgc gtgtgaagct cgggaatggc gctgaccgcg ccgtacgact 7740 ggtcttcgtg cctctgccag cagcgttcca gcgtgtcgta tataagctcc gcctgtctgt 7800 gtcccttgga gcgtattttt ccgcttccca cgtgaccgca ctcggggtcg agacagacgg 7860 cgtctttgag tccgtagagc ttgggggcga ggaaaatggc ttcggagctg tacgtgtcgc 7920 cgccgcaccg cttgcacttg atgtcgcagt cgcaggccca gtagagcgcc gggttctccg 7980 ggtcgaacgt gagccgtgtg tgcgggccct tgatgcggtg cgcgcccctc tccttcatgc 8040 gccgataccc cgactcggtg acgaagaggc tgtcggtgtc cccgtagagg gtctgggggt 8100 ccctttggag taggtgtgtc cccctatcgg gtccgtgcag gatttcgcac cactcgctga 8160 agaaggcgcg cgaccatccc aacacgaagc aagcgatttg cgtggcgtat ctcttatttt 8220 cgaccatttt gtctaaattt tctagatgta gaacggtcaa cgcttcgggg ggagcgtcga 8280 gcaaccgcat gggtttgaaa acggtctcgt tggcggtggc gtaatgggcg tggtcatcct 8340 cggctatata agggggcgcg tcgagcgctt cctgcagttg gtgatcgact tccgccaggg 8400 acgcttcgcg agcggtaagt gcgttttcga gcttactggt actctcgcct tcgctctctt 8460 ccgtttcctc ctcctcctgc tcttcgtttt cttcttctgc tgtccgcgtc gggggtttcc 8520 ggtcaaagta ctgacgcagt tgatgatgtc tgaaggcgtc gggggtgctt tcggggcaga 8580 cgaccgtgcc cgctagcgag tcgtggttga gcaatgtcac gtgacgcacg atttgagtgc 8640 cttcgtagat gccccgcctg tcgtcctcgg tgaggtcctg ttcgaagagg atgcgcgtgg 8700 tgtccatgtt ggtggcgaag gcgccgtaga gggcgttgct tagcattttc gagatggacc 8760 gcataacttc gtttttgtct cgatccgctt tctctttggc cgcgatgttt ttgctgacgt 8820 attcagcgca gacggttttc caagcgggga acacgatgtt catgtcgtcg tggagggccg 8880 tgaccgccca tcctcggttg tgcaacgtga ggatgtctat gacggtgatg acctcgtcgt 8940 acagcgcttc gttggtccac acgaggcgcc cccctcgcct ggagcaaatg ggaggcagcg 9000 tgtctaagtg ttccgcgggc ggaggataag cctcgacttt taggatgcac ggtttgattc 9060 ggtggtcgaa gtagctgatg ggtcgttcgg aactcatgag ctcgttgagc tcgtcgacgt 9120 gtttggcagt gaatttgggg tccaagggca tgccgtgggg catggggtgg gtgagggcgg 9180 aggcgtacat accgcatatg tcgaaaacgt atacgggatg tctgtacggg cctagcacgc 9240 tgggataaca ccgcccccct cgcaacgctt gccgtatgta tttgaacatg ggacggtgcg 9300 gggcgtatat ttcggcgacg tactcggggg agggaagcct cttttttctt tttttggtgt 9360 ttttgtttgc gcttgcgcgc tcgacgtcgt gtacgtcggc gtcgtgaggg ttccccttga 9420 tgtcgttttt cttgagcgcg acttttcttt tgtacttgga aggggggcgc tcgaccgtgc 9480 tcgcgcgttc ctgctcttcg acgtacgcgg tgaaggtaag ttgcttccaa aaggcgtggg 9540 tgttgctggg aatggtgggt cgcaggaaaa tgttaaagtc gccgtgcatg ccgagttcgt 9600 ctttgaagta cttgtcgtaa ctgtcgtaca aggtgagagc taatttctgc gtgaccctta 9660 cgtcttgcat gcagtactcg aggcacgcct gcacgatgtc gtacggacga cccgggtgct 9720 cctgtcgcca tagcgccttc tgttcggcga tgacagaagg atcctcccag tatctcgcga 9780 cggggaaccc gtcggagtct tgttcgtaac cgccgatgga gacgtgttcg ttgacggcct 9840 cgtaggggca ctggcctttg gagagctcta gggcatacgc ggcggcggct ttggagagtt 9900 tggctccgct ggtgaactgc aacgtgtcgc gtaccatgaa tcgcacgaag acggtgcggg 9960 agtcgctggg cgcgacgact cctcgcgccc acctgtgaag ccgcgtgggg tcctttttat 10020 cgtagttggg gttgggcagg tggaagagga cgtcgttaaa gaggaggcgg cctacgcgcg 10080 gcatgaagaa gcggtcgcac cgacacgcct gcggaaagac gtcgcgccgt tccacgagtt 10140 cggtggcgag caagagctcg tcgaacttgc agatgttgtg tccgagcacg acgatgtgaa 10200 cgttgttaaa gtcgtcgggc agtttgagag gttcggtagg tttgtcgaac agttcgtagg 10260 ggatgtcgta gcgggaaagg tatttgcctt cgctcatgag cttgctgacg tagtctccgt 10320 tggctcgaca gtacctgtcc acgagatccc tggcgaagtg ttgttgcaaa cgggaccggt 10380 aggcgcggaa ccgccgggca acgaagccgg gccgcgtgtc tagccagtag aagccctctt 10440 gtaaagtttt tacttccggg tcaacgagcg ccaggtccgt ggccgcgcgc acgacgtcgg 10500 gatcgcccgt gagcatgaaa cacaacataa aggggtgcat acgttttcct ttattttcga 10560 agacggtata cgtctcgatg tcgtaggtga gaaagagttg cttacagtgg ggttgctgag 10620 cggggcatgc gaaacgcacg tgttgccaca ggtcgcagcc gaatttttgg accgtgtggt 10680 agaaaaaggc ggaccgcctt tcgttgcacg tgtgattgcg cacccagtgg cgtccgcagg 10740 cgggacaggt ttgcacggcc gtgcggctga gtatccaaac ccagtcgccg ttctggtctt 10800 gggtgaccag catgggggga agaaccggat ctccgcgttc cgcgacgcgt ttgacgtgcg 10860 cgagacgccc tcggaagcgg acgacgccga cgtaagcggg tttgaagacg ggcagtcttt 10920 ccggggggtt ttcttggtag tacccgtagt ctattctatc gtatccgcgg cacctgcgcg 10980 tgcctgccgt aggaacttgt gtagtcgcaa gaattttttc atgcctttgt ggaacacggc 11040 cggttgcaga cggaacggta cgccgtcgat gtagtagacg ttggcgcgga tgggatctcc 11100 cttggccgcc ttgaagtggg tcttggcatc gttttggctg agcgtcttaa ccaacggcgt 11160 gtcttccggt ccgagcagag tgttttctag gggaggcggg gagtacgcgg cacgggaaga 11220 ggaggaggca gacgacgaga ggctcatcgc atgcgaggag gcgcgtgccg agcgacgagg 11280 agttcctgtt ctcggtctct caggacccga cgcaggttgt ccgtgatggc ctcgttggtg 11340 ctgatggcca cgatgccccg gaatttgatg cgaaaactga tgtcgatgga gtcgatgaac 11400 tcttcgctga gattgagttg tttgaggacc tcttcgatgt caccggaacg gtctctgtat 11460 tgaatatcgg agagaaataa gtgttggtct gcttcgtcca tgccttcgaa ctgccccgtg 11520 cgctctatca tcatgagaaa atcgcgcagg atgcggttcc acagggtttc gaaaatggtc 11580 gcggggttag attgttcgct ccagatgcgt ttgaaaatag gctctgcgtt gacgtcccac 11640 cccacgatca gtacctgcag ggtggtgacg tctacgtacc gccggaactc gcggttcgcg 11700 acgaagtgcg agtaaagata gtagagcgtg gaggcgacgt gttccgcgag gaaaaagtag 11760 agcacccact ttctaagaaa cgattcggtg acgagcgcca cgtcggcgtc tcgcgtttgt 11820 aggagcagtc ggtaaaagct atccgtgaat cggaacagct cgtgtctgcg ggcggcgccg 11880 gagagttcgt tttgcagggc gtcaatggct tccagcacgg tgcggatgac ttcgtcgagc 11940 agcgtttctt cttcctcttc ttcggggggc atttcttcct cctcttcttc tgggaaggct 12000 tcttcttcgg gaggggccgg gggcggaacg acgcgtctcc gtctgatcgt tcgcggaaga 12060 cgatcgacga attcgcgtac ggctctccgc ctgatgcgcc tgacttgcga ggcggtgatg 12120 gctcgccctt gtctgtccct gggtcgtagc ccgcgtctgt cggggacgcg tctgttgcgg 12180 aggacgatgg cgccgcccga gaagggatcg cctccgtccc cgccaccgcc ggcgcgcccc 12240 gcgtaccctc tgcccgaggc cagagtgcta acgaaacatc tcgccatggt ctccgcaggt 12300 tcctcggcgt tcactctcgg atcgagtgcc gcttgtcgga tcaaactctg cgtgtctaga 12360 tcggcgaacg ccgcttcgaa gagcgtgagc cagttttctt cggcgaacac atcggatcca 12420 ggtaagaacc tatacgtgtt ttccgtatta aaatgttcgc gcgcgtaaca gaataaatag 12480 tggcacagag cgactcgcaa acgccgtatc gcttccaata aggtcgcgtc gtctctcgac 12540 gccgcacgca tgataatgtc gttttgcaaa cctcccgcgc cgctacccga gaggtggctt 12600 cgcgcagcag cgagcgcggc gtccccgtcc ttcccgacgc ccacgacctc cccggagcgc 12660 gcgacgtcga accctctgcc tcgcagatga acgtgcgcca tgtcggaggc gattctgtcc 12720 ataagcacgg cgttgtgcat ctgctgaaag ctgtcgtgaa agttttcgag gtccaggaac 12780 ctcatgtacg cgcctacgtt gacggagtac gagcagtcgg tgaggcacgt ccagaaaagc 12840 ctcctgggtc gctgcgcggg cggactttcg tagccgagtt gcatgaagac gcgattttcg 12900 aaaaagtaat cgttgagcgc gcggtgcatg tactggtagc cgagcaggag gtgcggcgga 12960 ggcagtcggt gatacgggta tcccgcgacg ttgggtccgt tcgggctgag gtccctgagc 13020 tgcatcaatc tgtagtcgta gagtcggctg acgagaaaga cgctgttggg gtgtaccagc 13080 gccggctgat tcctgaccat gggaaagtcg gacgcgataa cgggttgaca aaatcgaacg 13140 gtgtttaagc tttgaccggt gagctccgcg aagactctgt aagcctgaaa ttgagccctg 13200 acgtttttag aggagggcct gttcccaacc gagagacgcc atgcacccgg tactccaaaa 13260 cgttcggaac gcgagcgcgg gcgccggcgg ggaaggacca caccgtcacc agcatcagca 13320 gcaacacgtg caacgccacc accagcaaca gcaacggcac cgacagcagc ttccgcaagc 13380 ggcgccgacg cgaaaccgcg tccgccctcc cacccctccg cagtacccgg cgcagcacgc 13440 gctgccctcg tccgcttccg cggacggctt cgcgtccgac ggcggcgagg acctaggtcc 13500 tcccgtttgc gggctggcgg cgggagcgga cgtagacgag attcgaatgc gaaagcggga 13560 aagcggtcgc cgcggagcga ttcccgagac gaatttgttt aaggcgagca gagaggcggt 13620 cccgcaaaac gattacgata gggaggcgat gtaccggtcg gggcaggcga tcagcgttaa 13680 ccggggtcgc gtgctgaccg ccgacgactt cgcgtacgat gagggacagg acccgtcgtt 13740 cacgccgggt gcgaatcact tgcgcgcggc cgagcttgaa acgcccggcc gagcagaccg 13800 cgttcgggaa gagctgcgca acagctgcca ccaaacgcgt ttgcgtacgg cgttgacccg 13860 tcccgagctg cccgcgggga tctactacct gtacgacttc gtgcagacgt acgtcgatca 13920 tcccgacggg cgagttaagt taaacccgca gctggtttta gtggcacagc acgcgggcaa 13980 taccatgctc gcccaacggt tgtggtcgat agcggaggat aagaacgcgt ggttgcgtga 14040 cctgatcgag atggcgtaca tgatcgtgaa cgatccttac ttgagcaccg agcagcaatt 14100 gtcggcgata tgcacgaccg tggtggagtt gagcatgaaa tacgctaaga tcgcggcgac 14160 gaacggttat ccgtcgatgg cgcagatggc taaggcgcaa gaattctttt accgcgtgat 14220 ggaagccgtg ttagacttgg gagttcaggt tggggtgtac aacaaccgtc cggtgcggtt 14280 ccgtcagaag cgcatcggcg agatccctca gatgtctgac gcggactaca tgttcggttt 14340 gacccaggcg ctcgagaaca ggcctcctca gggcgaactt ttcccgtcgg acgagggaga 14400 atccagcggc ggcgaggcgg aggaggacgg ctacgatgac gactactgag tcgtttatgc 14460 ttttggcccc tctgggtcgc gaggaagtgg cggacgcctt aagctcgtgc cggacgggcc 14520 gacggcgcgt tcgctttcgc catgctccgt actcgaaccg tttgcttaag ttgcaaacgg 14580 cgatggttcc gccgaagata gacggtactt cggaacgcgt ggccgaagtg gttaggggtt 14640 tggccgagca gggcgccatt tatccggacc agatgggggc catacattcc gatctgctaa 14700 accgcgcgta cacgtggaac tcgatgggag tgcaagagag catccaggcc ttggtgaacg 14760 acgtgattca cgggcagaac aaggcgctgc aagacgagct ttcccgtacc cgcgagatcg 14820 cgaacgcgtc gctcttgacc caatttttcg atagcattta caaaacggtg gacaggggcc 14880 agcgtaactt tgagggattt aaaaagctgc tgcggctttt cgttaacaac gtccctaacg 14940 cggaggtgta cggctcgggg ggttcgttca gcatgcagat caacctcgga ggcacgagcc 15000 agaacatcaa tttgactaac gcgttcgaaa acttaaagga catatgggga gcgcgttggg 15060 acgcggtaaa caatccccgg ataggagcgc tgctcacgcc caacacccgg gcgttactgt 15120 ttttcgtgag cgccttctac gattacgggg ctatggagcc cggcagttac ctggataaca 15180 ttctgagatt gtacaaagag gccattaggg cggacataga cgcggaggga gacgccgtga 15240 tggagttggg cgaagcggga gccaacctaa atctgcagtt caacgagtac aaggataccc 15300 tgaactacct gcttcagaac agacagtcgg tgccggatac ggcgccgctc gaattgagcg 15360 aagagcagga aatgcttttc cgctatctga tgcgtcaact aaggcaagcg ctgaaggaca 15420 ggattccttc ggacgtttcc gtgagcacga tggcccaata cgtggacccg cgcctttatc 15480 agacgaataa ggtgtttatc gaaaagcttc aaaactacct gttggccgcg cacgcccgca 15540 atccccatta ctttagatcg atcgtgatgg atcgagactg gcgtcctccc gccgggctct 15600 acacgggtaa ctacgtcata ccggaacgcc tgccgttcga ttattacgac ggcgacagcg 15660 tagtcggagg cccgtcgcga gacgagtact tcgaggagta tccctacgcg agcaaaccgt 15720 acaaaaagat gagcggtcgg gagagggccg actaccagga gcagctggat aacttaacca 15780 cgcagataga tcaggctttg ggcgtgcaat cggaggccgg atggttagcc gatcaccgct 15840 tgccccaagc gttcgatggt gctttgagtc tagcggatct ggcggcgatc ccgttacccg 15900 ccagtccctc cgatcaatcg atggcttcga gccggcgtag ctccctgtcg agcgtaggcg 15960 acatgatgtc gcgcctgaac ttatcggggc gagggggagc gggatttttt gcgagcctca 16020 gaccgacctt aggaagccct agaccttcta gcagcgccag acctagcgga ttagcgttgg 16080 gcctgagggg cacgggtcct gcgcgcgctt acagaacgcc tcgcgtcggc ggcatgtcag 16140 gctccggatt ggctagaagg gctttggcga cgcggtctat gcgcgacggg aaaaggctgc 16200 ggttttacta gcgtagatgc gtagaaacgg aagacgaaga aggaagaatg cgcgaggtgt 16260 aacgagggaa cttaccgcca tttctagagc gatgcttggt caccacgact ttggtttccg 16320 cgaagcggcg ttgaaccggg gttcctcttc ctccttcgcg acgtcgatgc cggccgctcc 16380 tcctccttcg cctgtgtcgg gcgtgccccc tgcgatggct cctcaacgct acccggcgac 16440 ggtgcacgga taccctccgg tgattccgcc actagactcg gccgcggacg acacgtccga 16500 actgttcgtt ccggtgcaga gggtgatggc gcctacgggt ggaaggaaca gcataaagta 16560 ccgcgattac gcgccctgtc aaaacactac caaattgttc tacgtggaca ataaactgag 16620 cgacatcgac acgttcaacc ccgaggcgaa tcacagcaat ttccgaacca ccgtgatcca 16680 taatcaggat cttgatccgg ctacggcggc gaccgagacc atccagctgg acaatcgatc 16740 ctgctggggc ggggaattga aaaccgccgt gaaaaccaat tgcccgaacg tcagttcgtt 16800 tttccaaagc aatacggtgc gagtaaagtt gatgagttac cgcgatccgg tgcctccagg 16860 cacggcagct cccacgagcc ccgagcccta cgctccagcc ggtgctcagt acaagtggta 16920 cgacctaacg attcccgaag gcaattacgc cttacacgag atcatcgacc tgctgaacga 16980 gggagtcgtg caaatttatt taaaagaggg acggcaaaac aacgtgctgc gctccgacat 17040 cggcgtaaaa ttcgatacgc gttactttaa cctgcttcag gacccggtga cgggtctggt 17100 cacgccgggc acgtacgtgt ataagggcta ccatccggac gtggtactgc tgcccggatg 17160 cgcggtagac tttacttaca gtcgactaag tttaatgtta ggcatagcga aacgcgaacc 17220 gtattcgaaa gggtttctca ttacttacga agatctcgag gggggtaacg tgcccgcact 17280 gctagacgtg gaatccgcgc agatgaccgg cgtcgatcaa gacgtgatcg agctggcgga 17340 cgcgaagccc ttgttaaagg actcgaaggg cgtgtcctat aacgtcattt acgactctaa 17400 caaccgtccc gtgaccgctt atcggtcttg gcttatcgcg tacaaccaat cgggttctac 17460 ggcaaacgaa acgaccttat tgaccgttcc cgacgtgggc ggaggcatag gagctatgta 17520 cacatctatg ccggatacgt ttgtggcgcc taccgggttc aaggaagata acacgacgaa 17580 ccttgccccg gtggtgggca tgaacctgtt ccccgccctc aataaagttt attaccaggg 17640 cgcgtccact tacgtgcaac aactggaaaa ttcctgtcag tcggccacgg ccgcctttaa 17700 ccggtttcca gagaacgaaa ttcttaagca agctccccct attaacgtgt cagccgtgtg 17760 tgataaccaa cccgccgtcg ttcagcaggg ggtgctcccg ttgaagaatt ctttggccgg 17820 cttgcaacgt gtgctcatta cggacgacca acgtcgacct ataccctacg tatataagtc 17880 cctggctacg gttcaacctc gcgtgttgag cagctccact ttacaataaa acactcaaaa 17940 atccaaaatc gtcgccgatc gctgaaaacc atgtccattc tgatctctcc caacgacaac 18000 agaggttggg gcatgcgtcg ccgctcgtct atgcgtgggg tcgggattcg tcgtcgacgg 18060 ttgacgctgc gtaccctgct cgggctgggt accagcagcc gacggagaag cggcggtcgg 18120 tccagcagac gccgttcccg ccccgcttct accactactc gcgtaatgct ggtacgtacc 18180 agcccgaaga aggagacgtt aacgcgcgag gtcgactgga aaacgcagat ggaaggagga 18240 ggaatgaaaa attgaaggag gaggaggacg aggacccgcc gttcattttt cttgactgtt 18300 gttttaacac tcgtaatcgg ctgttacgcg agaggcttgt aaataaatgg agagaaaatt 18360 tgcgcaacgg aatgagaaaa gactactgat ttgcctgtac cgtcgctgta ctacgcagga 18420 agcgtcgcca acatcatgcc ggcagtagtg cttacgggcg ggcgaacagc tagagctaat 18480 cggagggcgt caaccgcttc ccgccgaaga ctccctgctt ccaagctacg ggccacccgt 18540 cgaacgcgcc gctcagtcaa cggtagtaag aaaaagcgtt cgccgaacgt ggctgtggcg 18600 gtaccgtcgc ccactacggc ctccgctgcc gagcgagccg cgctgcagaa cttagctacg 18660 cggttacagc gagggcagtt tacggcttgg cgttcggcga actaccccgc ccctgcggcc 18720 tccgaggccg ccctggccgc tgctcaaagc ggcgcacccg ctacggccag ggacatgacg 18780 accggaacga ccgctacggc ggttcccgtt tcaggtgccg ggatcgcgtg ttcaacgagg 18840 agaaggagaa acggtgttag cggcggtaga acgcgtcgcc ggggaaaaca gctgaaaggg 18900 ggaattttac ccgctctcat acccatcatc gcggccgcta tcggggcgat tcccggaatc 18960 gccggcacgg ccgtcggcat tgccagtttg aaagaacaac agagacagtt caataagatg 19020 tacaatcaga agtaagagag agaaaagaaa aaaaccgcgg gtgccggtgc tgtctaaaag 19080 gccgttttca ataaaaattt ttatagaacc gctgtcgccg ccgtcgtttg ccgtttgtcg 19140 tcgtcgtcgt cgtgagcagt tgtcatggat tacgccgcgc tttctccgca cgtcggtggt 19200 tgggccctgc gagacggata cctcggagac tctagcctgc gagggggtgc tatcaattgg 19260 tcgaacgtgg gctctcgtct ctcgagcgct ctcggcacgg ccgggcgatg gatgtttaac 19320 cagggaaatc gctttctcaa ttccaacacg ttcggccaga tcaagcaggg ctttaaggac 19380 agcggagtga tccgtaacgt cgcaaacctg gccggggaaa ccctaggcgc tcttaccgac 19440 ataggtcggt tgaaaataca acaagatcta gagaagcttc gccgtaaagc cctgggagag 19500 gacggacccg ccactcaagc cgagttacag gcattgatcc aggcgttgca agcccaagtc 19560 gccgcggggg aatccgttcc cactaccgct cctacggtag ggccttcggc tcccgccgaa 19620 gtgcctcgac cgacaacacg ccccatcccc gagatggtca cggaggtgag gcctcccgtg 19680 acgtcctcgg ctcccgcggt gcccgaaacg gaccggccga cgaccctcga aatgcgtcct 19740 ccgcccgcga agaggaggag aaagaggcca gcccccggat cgtggcgtac tcgcctaaac 19800 agcctttcgg gaaacggggt cgcagttagc cgccgtcgca tgtgttattg acctctcgat 19860 aaaaaagcgt ggagatgaaa aaggcattaa atcgtatcta tcggattacc aaaaatttag 19920 catgtcgccg tttttttagg tcgatccgcc gtcggctgcc gctatggccg cgtttacacc 19980 cgacctgacc accgcgaccc cgcggctgca gtactttcac atcgccggtc cgagcacccg 20040 ggaatacctg tccgaggacc ttcagcagtt tatcgcggcg actggaagct actttgaact 20100 gaaaaacaag ttcaggcaga cggtagtcgc gcccactcgc aatgtcacca ccgaaaaggc 20160 tcagaggctt cagatcaggt tttacccgac gcagaccgac gacacgccca acagttaccg 20220 cgtgcggtac agtttaaacg tgggcgacag ttgggttctg gacatgggag ccacctactt 20280 cgacatcaag ggcgttctag acagaggacc ttcttttaaa ccatacggag gaaccgcata 20340 caatcccctc gcgccccgcg aagccttttt caacaattgg cttgagtcag agccgaacaa 20400 gaccgtcatt acggggttaa tgaccacccc ctaccgaaac gaccagggga acaagaccaa 20460 tactgctgac gtagttaata gcgtgtcggg agtttatccg aaccccaatc tcggaccctg 20520 catcagcgag atgggagcac tggatgatgc aactgctgac cttgtcggct ttgcgggacg 20580 gtttgcgaaa gttacaaacg agaatacgcg tttggcatac ggagcgtatg tgaaaccgtt 20640 gaaaaacgat ggttcgcagt cattaaaccc cactccttac tgggtaatgg acaagactga 20700 cgccaagtat ctgggcgtga tggccgtgga agaattcagc accagtctca cctatccgga 20760 cagcttgtta atccctccgc cctcggacta ctcaaccgtt aacacgggcg caatgaaagc 20820 aaaccgtcct aactacatcg gtttcagaga taacttcatt aatttgctat atcatgatac 20880 gggcgtgtgc tcaggtactc tcaattccga acgctccgga atgaacgttg tcgtagaact 20940 tcaggacaga aataccgagc taagctatca atacatgtta gctgacatga tgtcgcggca 21000 tcactactac gctctgtgga accaggcggt agatcagtac gaccacgacg ttagggtgtt 21060 caataacgat ggctacgaag aaggagtacc aacgtacgcg ttctctcccg agggcacggg 21120 atatggcgct aatcctaatt ctaacctgtc gtttgctgac gtgaaagtgc atcaaaacgg 21180 ggcgggcgac aaaggaaacg agattaacac gggaggtgca acttactcta catacttaac 21240 cgtgggaagt gtgccatctt atgaaatcga tcttgctgct tctcagcggc gaaacttcat 21300 agtgtccaat attgccgatt acctgcccga taagtacaag tacaatattg atggttttaa 21360 tcccgagacg ggtaacgtag accctactac atacgaatac atgaaccgca gggttccact 21420 taccaatgta gtagatctat ttacaactat cggcgccaga tggtctatag accagatgga 21480 caacgtgaac ccattcaatc accatagaaa ctggggtctg aagtataggt cgcaattgct 21540 cggtaacagt agatattgca gattccacat tcaggttcca cagaaatatt ttgctattaa 21600 gaacctgttg cttctccccg gaacttacac ctatgaatgg gtgctgagaa aggaccccaa 21660 tatgattcta cagtccagct tggggaacga tctgagagcc gacggagcct ctattgtata 21720 ctctgaggtg aacctaatgg cgaactttat gcctatggat cacaatacta gtaaccagtt 21780 ggaattgatg atgagaaacg ctaccaacga tcaaacgttc gcagactacc tgggagctaa 21840 gaatgcgctt taccaggttc cagccggctc cactgcactt actattaata ttccagcgcg 21900 tacatgggag gggatgaggg gttggtcttt tacacgtgtc aaagcctctg agacacctca 21960 aatcggtgct caatatgata tcaacttcaa gtactccggt tctattccgt actcggatgg 22020 taccttttat ctcacgcaca cgttcaggaa catgagtgta ttgttcgata cgtctatcaa 22080 ctggcccggc aatgatcggc ttctggcacc caatctattc gaaatcaaac gtaacgtcag 22140 cataaatgcc gaaggattca ccatgtccca gtgcaatatc actaaggact ggtatcttat 22200 ccagatggcg accaactaca attatgtatt taacggctac aggttctggc ccgacaggca 22260 atatttccac tacgattttc tgcgaaactt tgaccccatg acctgtcagg gacctaactt 22320 ccaagatcaa actttatttg atctcatgag ttacgatcca gtgcagactc ccggagccgt 22380 tcagaccaac caagacgcta tccgtaacaa ttccggttac acggctcctc gcagctggcc 22440 ggtatacgga gcacaacagg gagaatcgtg gcccgcgaat tggccgtatc cgcttatcgg 22500 tgacagctca ctcaatccta cccagatagt aaattacaaa aagtttttgt gtgataatta 22560 cctgtggacc gttccgttca gctcggactt catgtacatg ggtgaattga ccgacttagg 22620 gcaaaatccc atgtatacta acaactctca cagcatggtc ataaatttcg aactggatcc 22680 tatggacgaa aatacttatg tatatatgct ttatggtgtg ttcgatatgg taagagttaa 22740 tcaacccgaa cggaacgtgc tcgccatggc ttacttccgt acgcctttcg ctacaggcaa 22800 cgcggtgtaa aaaaactgat taaaaaatgt ccgggaccac ggaaagccag ctgaatcaac 22860 tagtcggggc catgcacctc cgccatcgat tcctgggcgt cttcgacaaa acttttcccg 22920 ggttcctcga tcccaaccgc cctgcctcgg ccatcgtgaa taccgggtct agagcgaccg 22980 gaggtatgca ctggatagct ttcgcgttcg atccgatcgc gcgaaaatgc tacatgttcg 23040 atcccttcgg ttggtcggat cgggaactgt ggaatttgta caaagtgaaa tatgacgctt 23100 ttctcagaag aactggatta agacaacctg ataaatgttt tgaactcgta cgttccgtcg 23160 aagcggttca atgtccgtgt tcggcggcgt gcggactttt tagcgctctt tttattgctt 23220 cgtttgatcg ctaccacacc cgtccgatgg atggtaatcc catcatcgat acggtcgtcg 23280 gcgtaaaaca ctccgacatg tacaaacccg aattccagag cattttgcac cggaaccagg 23340 agcgaatgta tttctggttt atgaaaaaca actctttttt tcgcgcacac gaatcggaac 23400 tgaaacgcga aacggccatc aacagcgttc ccgaaaacca ttaagacatt acactcttgt 23460 aaatttctat atctgtattt tttattttcg cattcgcgac aataaactac atttattttc 23520 gagaccgcgt gcgattattg ctctcgtccg cgcccgttat gcgaacgggt cgcctgtttc 23580 aacgggttta accggagcga tgacctcggt tttaaacccg aaacgctcgt gccaacggaa 23640 ttcgggaacg tgagtcggcg cttgaacgcc catggcgttc aacactaatt ccgtggcaaa 23700 aacgtaagcg tggcgtagat ccatcgcgga aagccgccaa gcgcaggttt tatcggaggt 23760 cttgcgaccg gagcgagagg tgggaccatc cgtgcccgag ggagattgcg gattgcagca 23820 cgtgtacacc atggtatgcg ggtacttctt gtgcgccttc atatcgggac gcgagcgagc 23880 ggtatcggaa gcgatatcgt cggtgccgct cagcttgtat gggatcatgc gacaaacctg 23940 acgcccgctg atgggacctt gaacggcgta attacaatta cagttggtcg atatcagaat 24000 atgctcttcg gcacggcgtt tttccgcatt tggatagaga gccttggtcc actctaagtc 24060 gtgtctcatg gcgctcaccg ctttcgcggc atccgagaag gccatgccgc agcttccaat 24120 cacgtgaggg tacgggaaac cagagtggtc ggcgtccctg gagcacacgg cgttacggtc 24180 gaaccgcaag acgaccactt gtttaccgaa gcggttcttc tcgatttgac cgttttgctc 24240 cgcaatggct ctctttcccg cttcgctagt agggttgagc tcgacggtgc gcggcttcaa 24300 gaacatggca ttaccatgca agcatttggg ggtggtgttg gtggtggctt cccatccgtg 24360 tcgccatacg tgcgctcctt ccggaacgaa cttcggttcg agctccgcca ttccgtacac 24420 catggccgcc agaaaacggc caacttggtt gtggaaggag tcgaaactcg agaaggtaag 24480 tctaaactca ggatgcctct tgcgcacgaa tgtacctcca atcctggtcc aaatggcatc 24540 gtcggggcga atagtggaac cctgccaccg gacatcgagt cggtcgcaaa tcgttgtcag 24600 aatcttcatg gccttctgcg caccgtactc tacgggatcg cttaacggcc tttctaattc 24660 ttcatccgat tcaacgtcga gttcggattt cttttcgtcg acttcttccc gttcctgcgt 24720 ttccttgcgg gcttttttgc tgggcgcggc accccgagac ttagcgctcc tcttcttttt 24780 cgagtccacc atcgctggtt cagcgttctc atgatcgtcg ctagacgcca tctttttgga 24840 ggtagtttta ggcgcggaat cctttaccat gccgctacga tcggtttctc cttcttcctc 24900 ttcttcgtcc tcacccggtg agctctcttc agaactggag aatacttcct cgacttccag 24960 cctcctcttt tgtaccttga cgggatgacc tattggttga acgaaaccgt tttacgacga 25020 cgtcacagac ccctcccata gcccccagcg cggtataaaa gtagatttcc gggtcggtac 25080 gaccactctt atgccatcga gggaggctgc aggagcccgt acctcggcga tccgacggtg 25140 gaagtgtctc cttcggtagc ggtgcttcct gcagcggcct cgtctttggt tggctccaat 25200 ccgattccga gtcgtccgaa ttgggcatcg gagtcttttc cctggctccg aatacctgtc 25260 gaatctcctc gtcgtcttcg tccgagctca ctacccaaag atcgcggtct gctgcctccg 25320 ccatcacggt tcttcctccg gctcctccta gagagttcag atcgagggaa ggagagaaaa 25380 cggcatcatg caaacggctc gcgaagggaa ggccacggtc attctgaatc tcgtcatcaa 25440 taccgcgaag gaaaaaatcg gttggcagcg cttcgtggac gcgatccaac ggtacgtggc 25500 cgaagcttat ggtgctttct tggcgctcga cccagaggcg gcaccaccta gacgcgaatc 25560 cgaccacgcg gtgcgagttc ttatcgatgc tttaggggag gagagagcag ttttagctgc 25620 ttatcgcgtg gcggaggagt tattagaaga taaggagatg ccaaaagaaa gccaagacaa 25680 agaacctttg tctgaaaatg aggaacttgc gcattctttc gcggaaagta ccgtttctcc 25740 gcattctttt gcggaaagta ccgcgacggg tgatataaac gtacccgatg acgagagttc 25800 agtcgcctca caaacttact cgggagagga acgggacgag tctgagtgcg actcggacga 25860 agaagaaaac gacaaggagg cgaacgaagt cgaagcagac gaggcagaag agcaggagga 25920 ggaggaggag gaggaggagg aggacgaaga agatgactct agcccctcca ccgtgctgga 25980 aaacgagcga atacaagaaa acggggaacc cgaccacaat cccaatggag acgagggagg 26040 agctgattct gatgcggata gcggatacta ttcggcggac gcgggacgat tgggatcgtt 26100 actttcggac tcagaacacg aaaacgacac cactgaacgg caagtggaag tgggtatcga 26160 ggacgagtct cctactggtc accatccact tgggactcct gattccgatc gcgatcatca 26220 ggagcaagaa acttctatag agacgaaaca tgtagatccc gtttctactt ttaaaaagtg 26280 cttcgaacgc caagctgctc tgcttaccgg cgcccttaag attccttgcc tgctgaggac 26340 gccgacgaac ccgtctccgt atccgcttgt gcagtaccag ctagagagat tcgtattcaa 26400 tcccgaccca agggtccctc ccgaacaccg cgaagtgcgc tacaatttct accctccttt 26460 catgcgtcca aaagctatcg ctaattatca cattttcgcc gttaccgccc ccatccctgc 26520 tagctgcaaa gcaaacagaa gcgggagcaa attactagag tcctgccgac aaacggcaac 26580 gtttaagcgc ttacctcggt ggcggatcaa cgttcaattc gatgacgggc tcggagaaga 26640 ggtgattcct gtaacagagc tatcagatgc aaagttagtt cctttgcaag aagacatttc 26700 gagattgcag tgggccaaaa tgagaggtga acatattcgg ttctttagct atccctcttt 26760 acacatgcct cctaaaattt cacgcatgct catggaaacc ttgctacaac catttgccga 26820 tgaaaacgat aaaggcgaag gaccaaaacc gtgcgtgagt gatgaggaac tgcgttatat 26880 cgtagatccg gacggcaaaa tgcgcgggga agaactctat aaagctatgc agcggagacg 26940 aaccattgtt accatggcag ttcgttttac cgcgcttctg gaactcatgg aacgcgtcct 27000 cagggagcct tcttcggtca aaaaagcaca agaggtgctc catcatactt tacaccacgg 27060 atttgtcgct caagtacgcg aaacggctaa agttaatctg agtaattacg cgacgtacca 27120 cggaatcacc tataacgacc ctctaaacaa ctgcactgtc gccaagctct ttgaaggtcg 27180 ggacaaggaa gattatgtgc tcgacagcat ttacttattt ttagtcttga attggcaaac 27240 cgcgatgggc atgtggcaac aagccataga cgacaccaca attcaaattt acaccgaagc 27300 ttttacgcgc caacggagag ccatttacgc gctgactagc gtcaccgagg tgtccaaagc 27360 catagtcgat cttctcatgg acggggaccg gttaaccgaa gaaatgcgca aagctctgcc 27420 taactttatt acgcagagcc agttgtctga ctttagacac ttcctaacgg aaagatctaa 27480 cgttcctacg atggcagcac ccttctatcc ctctgacttt attccgctag ccttccgcca 27540 aagcgcacct ctcttgtggg atcacgtcta tctgcttcaa gcggcttact tcctcatgaa 27600 tcacggcgga tacctatggg aaccctccga gagcgatgcg gaatctccgc aatttagagc 27660 ttactgtccc tgcaatctgt gcagccctca tcgaatgtta gcagacaacg tggctctgca 27720 caacgaagtc ctcgccatcg gtacctttga gatcagaagc gcggatggta aaacttttaa 27780 acttaccccg gaactatggg caaatgccta cctagataaa ttcgtggccg atgacttcca 27840 cccgtttacc gtttttcact attccgaaaa ccgctcatct tttggcaaaa actataccgc 27900 ttgcgtcacg gagagccccg aaatcctatc gctgattcgc cagatacagg cctccaggga 27960 ggagtttctg ctcacccggg gcaaaggagt ctacaaagac ccacaaaccg gggaaacgct 28020 agccacttcg gcggcgagcg tcggcgaggc tcgacctgga gctgccaggg ggtgcgtctt 28080 taccgtccgc tcccaccggt gccagcggag gaactccagc gccgcaaaag cctcctaggg 28140 ctatacgggc tgccgatcag atcatctccg gagactcgcg acataccagg cgcgactacg 28200 gggaaccgtc tttatcaatt ccatccgact ggcgatatgg ctcagagaat gctcgacgag 28260 aagaccagag taaccgaaga gccacccccg ccgtcggaag agaccgacgc tcccttccat 28320 acagacgggc tgtacgcggc acgggaagcg atcgagtccg acggcgaaac tctaggaagc 28380 caggatacgg aggaggacct gagtaccatc tcggaggagg aggaagagga gacttacgag 28440 gaagcggaga aggagaatat acctccgtcg aggggggcat ctcgaaaacg ccgctccgca 28500 tcctccctag agaagactct cccgagacct ccgctgagaa agaaatcccg caataacgct 28560 gacgagggta atgcccaagg gcttagaaaa cgtcgcccta gaggtaacta tcgcagctgg 28620 gtcagacacc gggtggctat ctgtcaagct ctacgcgatg cggtattcga ccgcaaaatg 28680 gctgctgaaa ttctcaaaag agtgcacggg ttgttcgtgc ctcctactgt cttaggctat 28740 tacgctcgaa aactcttaga tcttaccgac gaagactctg atcgtcggtc tccctttttt 28800 atgttcttag tgtccgacga tgccgtaatc gacgaagagc cggttccacc gcctctacct 28860 cgtaaacggc gccgtagggc gaataaggag gaaccgaacg cctctgaaac acagcttccc 28920 gaacccgtat ccccagcagt atctgatttg aaagccgaaa tattgaatct gctcgttgaa 28980 atcgaaagtt tcgtccgtaa aaacccgtcg cgccgcgtct cgattcgcaa ccgtacccgc 29040 gagagcatta cgcgccagct gcactacatg aaaagtgaac aaaaactcac caagcttaag 29100 acagatgcgg aaaaaatcct acacctgtgg agatcccttt cataatcccg cttcttttat 29160 agccctccgc tgcgcttcaa catcatgaac ctgctcgacg ccacgcctac cgagtacgtg 29220 tggaaataca acccgctctc aggaattccc gccggcgcgc aacagaatta cggagcgacc 29280 ataaactggg tggtgcccgg aggcaatagt ttcgcctacg cagcggatga gataaggcga 29340 aacaccctga gccccgccgt cacgcgagcg ataaccgcgc gcttcgaagc tgaatcagac 29400 cagcaaccct tcgccaaccc ccgggaaacc gcctacatca ccgctaacgt actggactct 29460 ggctttccaa agtccgcggt ttatcccgta gatccttccg gagtacaaaa agttcagctc 29520 gctggaggcg ccgaggggcg tatgcagctc tcaggcggcc tcactgaagg tcgagtgcaa 29580 ctttcgggtg gtgtattagg acacgtgcct cgccgcagtc ggcgaggcgc cagacctcct 29640 cgatggtgcg gaacagccct cgcgggaaac ggactaccag aagacgccga aatggtttcc 29700 gacacctaca agtatttcct ccgcactcaa ggacccagcc aagtggtctc agaaccgggc 29760 gtatactctc aacggcagtt tatgacaacc tttttacccg ctgttgtccc acatcccttt 29820 gacagtccta atcctaacga ttttcctgct caatatagtg ccatctacaa gggcacaaac 29880 gcttatgagg atgtattttg ggactggtaa gccggtcttc acacttaccc gctgctgtcg 29940 atgctcagtt tcgcaataaa gttcctccaa ttcacgttcg ttaaacggtt cccgtctcgt 30000 cattattacc cgttcgccgc cgtcgctcgc tattcgcgct ctaaaacgtc ttttgagcca 30060 aaagacgtaa ccggggttca ggggttgcgt aaatctaacg atcgcctgat cgttgatttt 30120 caaccaatat tttttaggag catccacttg cgcctccgac atggcgaaat cgactccttt 30180 cgcgttctcc atggggcagc actccagccg aaaacgtccc gcggacagcg aaaacacgca 30240 aaatgcatca aaagtcgcca aaacgcagac ttctgccacg cgcgccggtg tcgacggaaa 30300 tgacgaccta aacctggtgt accccttttg gctccaaaac agcacttcgg ggggcggagg 30360 aggaggaagc ggcggaaacc cctccctaaa cccccctttt atcgacccta acggacccct 30420 ctatgttcaa aacagtctcc tttatgtcaa aactactgca cctatcgagg ttgaaaacaa 30480 gtcactcgcc ctagcttatg actcctcact ggctgtggac gctcaaaatc agctgcaagt 30540 gaaggtagat accgagggtc ccatcaggat ttccccagat ggactcgata ttgctgtcga 30600 cccatcgacg ttggaggttg atgacgaatg ggaattggca gttaaattag atccaaacgg 30660 acctcttacc gcttcatcag cgggaatcaa tataaacgta gacgataccc ttcttataga 30720 agatgacgat gctaatcaag caaaggagtt aggcgttcat ctcaatccta acgggcctat 30780 tactgccgac cgagacggat tagacttaga aattgattca caaaccatgg ttgtcaagga 30840 cagcggaacg tccggcggcg tactcggtgt attattaaaa ccgagtggag gactgcagtc 30900 aagcattcaa ggtatcggag tagcggtagc tgatacgtta accatcacga gtaacaccgt 30960 agaggtaaaa actgacccca atgggtcaat aagctactct gctaacggca ttgccgtaaa 31020 acccgatccc tcaggaccgc tcacttcctc tggtaccgga ctatctgtgg ttacagcagc 31080 cgagggttca atccaatctt ccaatgccgg tttagcggtt aaaaccgatc caagcggacc 31140 gattaccagc ggttcgaatg gcttaaacct gtcatataat gcatcagatt ttactgtgtc 31200 acaaggagtg ctaaacatta ttagaaatcc cagcaccctc cccgatgctt atctggagtc 31260 cggaaccaac tatttgaata attttacggc acaagccgag aattctagtg tattcaaatt 31320 taactgtgcc tatttcctac agtcatggta ttccaatggc ctggtaactt cctctctcta 31380 tttaaagata gacagagctc agttttcaaa tatgccgaca ggtcagtccg cggaaaacgc 31440 cagatatttt accttctggg tacctaccta tgagtctctc aacctgtcac gggtttccac 31500 ccccacgatc acgcctaaca ccgttcagtg gggcgcattt tcgccggccc aaaactgctc 31560 aggtaatccc gcctttcagt acaatctcac gcaaccgcca agcatctatt ttgagcccaa 31620 atcgggttcc gtgcaaactt tccaacccgt attgacagga gcttggaata ccgacaccta 31680 caacccagga accgttcaag tctgcatact gcctcaaacc gttgtgggag gccagtcgac 31740 ctttgttaac atgacatgtt ataacttccg gtgtcaaaat cctggaatat tcaaggttgc 31800 tgctagtaac ggcacattca ctatcggacc cattttctac tcctgcccaa ccaatgaatt 31860 aacacgaccc acataataac ttacacgtgt aaatcaataa aaaagaatcg atgtaaaatc 31920 tttattgatt taatgattca ttgtcgcgtc ccattcggca cattgccagt aatacagaca 31980 attgacagcc tccacgaacc acgcctcact aattaaatgc accttctgtt caggaaaatc 32040 cacctgcatt cttcggcagt agtgacaagg gagtgccccc atttgtccga gcgttgctat 32100 ggcttccacg aaaatattaa ctttattggg gggaacgtaa attaccattc ccagaactgc 32160 accagaagcc ggtgaaggac aaaacaccgg ataaaaatca gtaaacatgg ctgcagccca 32220 agccgccaca aaataaactc cctcaagcgt gggatcatct tctaagcggc tccacaaata 32280 aactcccaga cctgtcccgt cgggctgaca acacaccccc atgggaactc ttttcatatc 32340 gaatccgtga caatcacatt tctcagtaat caactcaaaa tcttctaaac acaagaactg 32400 ataggcatgt gccgcgggaa caaatcccac gttgtttacg gcggcgggaa catcttgtgc 32460 ctgttcagca ccatcagcat ctgctagcgc tctctcggcc atctgtcaca cttgaacgtc 32520 ctcgatttcg gcgtacgggt gactgtcctc ggttaacccc tcgctaaaaa cgaccaatgg 32580 aacgggaacg ggaggagtag gatcggatgg tagccgatac aacacggagc gcaatcccgc 32640 ttcccctgaa tacagggggt tactgctaac tgtccaggta ggagatcttc gtggaaagct 32700 cgacagtcta cgcagccacg aatgtcttcg agatggtgtt ctattggacg cagtacgctg 32760 ccttcggtcc cgccttcccg tagcacacca aaaaatcagc aataaactca cggtagttgc 32820 tgtcaaaaag gtgcaacatc caaacacggt aaaaacaaag gtcagttcgg atatacggtg 32880 actcaggaac agatagctga caaccacgga agcgcagtaa gaacatacta ctgctgaaat 32940 gcagaacagg atacatagtc ttctgatccg agcgtcgctc ggtacctaac aataaaagaa 33000 aggcgatttt ctatcacctt ctactctagc acgtcttttt gacatgcaat tgcgtaagta 33060 actgagttgt gcaatataag ctagtggcaa cggttgcttg aggggacacc ccgggcatgc 33120 gctctcagga tgatgatcgc aataagtacc atgcacatgc aatacccatt cctccacctc 33180 tctataggta gcccgtctaa aggcgggaat gtaacacaac cccttagaaa caataggcca 33240 aacagaaggg tcggccatct ccctcaacat tgatcgtgtc attaacggtt gagtcacggg 33300 actaccactg ccgctcgttt ttaaaataac agtagctaca acattggtac ggcatgcgcc 33360 cagaaaaagg acaggatagg gcaagaacgg taactccctt ggcacctttg tcgtagacac 33420 ctcgtaacta acagtttgta acgcgaactt aaaagctaaa ctgttatcta acagaatccc 33480 tgatcgcaaa aatacaaaat cggacttgtg tctcctgata atccacttgc gatgcctatc 33540 taccaactcg gtcttggcag caaaaagcct tctcgaaaga acccgtaaaa actcctcctt 33600 ctgatcggga tccagttgca atcttgcttg aacccttaca atcacgttca agatgatcct 33660 cggccccgac tttaccctca catactttgc acatctacaa ctgactacgg aatacaggga 33720 ctcgtgccac agataatgtt gtggcttttc gaagaggtcc atgctcatga ttgtccgatt 33780 cgtgtgaaat tccatcatcc gttctaggga ctcctgagtc gcaccctgtc tcatcagtag 33840 actaaaccat atcactccgc actcattttg aaatccgcag cccctggtaa gggctaccac 33900 ctcctgtaga attacaagga tgcaagatga aatcagaata tagagaagaa tcagcgagcg 33960 catgcagcac atgcataagc acttccatga aactcaccga ctcagtacac agaggcatga 34020 tgtcgagact gatgattttt tattgatcaa gtaagtccac ctgaaacata ttaaacaaca 34080 tcataataaa tatcgcggta acatttacaa cggtgatggc agaatatata catcacgaca 34140 catcccgtga aacacgcagc ggctacaccg cccgtcacaa ccgcgatacg cgtaactcta 34200 tcttcccaaa attcacggat cagagtgtaa aactctcctc cgagatgagc aatggtgggg 34260 tgcccttcgg ttttctgctc cgagtacgta acagcgtcac catcatggct acaaatgcaa 34320 gtcctacgac aattaaaaaa agagcttccg tcgaaacacc cgtcgaacct tcttccgatg 34380 gaatctggac tggtgcttga agtgcctcca tccgaggaaa gggttccgat gcgtttagag 34440 ttatcattat cgcgtctgcg ttcgtagccg tctcgttcca accatattcg gaagtcggag 34500 taggatctga tgactcccaa accgaggtag cgttcaccga agtatagttc gcaataggct 34560 ccgctgtcgt aaccttctcg acaacacggt ttaccggtga tgaaaccgtt actttcaatt 34620 ttataacttc gaattgaagg aagaattcca gatcataaac atcttcagca aagaaagtca 34680 cgttaaactc tttcttcatt cgattcagat ggaaaacaca ctgagaactc gttacacacg 34740 tatcccaata actcactagt ctatgagtcc ccaccctgtc agtcagatga acactcatca 34800 accaaaacgg tgacctactc aaatctatct gtaacttgtg tccgaccgtt acattcatat 34860 caggtaattt actgacaatc ataccggcgg ttgtattctt atcgaccaag catcctgccg 34920 tcggcggaac gtttatatgg gcacctccat tggggtttaa cattgtacgg tattgcataa 34980 tcggaaaatg ctccttagta tacatacaat agtgagacgg gcgtccgaga ggatccggtg 35040 tgcgtgtaca atacatagtg cgaacagaac ccccggtcat aatttgtcta tttgctaagg 35100 gagcaacaga atcggatacc tggtaaaatc cctgaggcgt aatgacgtga actaaatata 35160 tacgggcatt cggatttaag tgatggatcc acagagaacc ttccgccaac tcggtatacg 35220 ctatctcggt ggttacatcg tatttcccat atcttaaagt ctgcacacta tagcctccca 35280 tatatacgct gctatcatgt gaagcaacta ctataatcat aatagaagaa acaaagtcct 35340 tcttaaaatt tggggaatac gacatttccg tcatatactt tatgctgtaa tggttgaagt 35400 aactgtcata tctataatca cgactaagtg tatacccatt ccaagtactc cttaccggtc 35460 ctaccctcga cccttcctgt aatacatgga acacaggcaa acctgtcttt acatccaggg 35520 ttttcaaaaa ctttggaaca atcattcgat gatagcatcc acttaaacta aacgtattac 35580 cccaggaaaa ttcctcacat actgtcgctc cccaaacatt ttctccgcat actcttccgg 35640 accatccacc gtgtatacca cagcttctac cgcccttatc ttccaccatt aaatcttccg 35700 aagccgtgta cccaccagtg atccacgaat actgagtctt tagaacgacc acataatccg 35760 catccaactt atctaatcca ccgcttacaa atgtcggttc tagcccgact attctgatac 35820 accgatggcc cttgctcatt tgtgaatact gtctacatat cgtcgagcaa gcatgagctc 35880 ctgccgagtg acctatacag tgcaattttt tagcgtcggg gatatctacc agaaacttcc 35940 taaaatccgc atgtaaacca tccgtatcat agtacaaccg atagtccacc aacacaacac 36000 ctacggcggg agtcatcttc tgatgaaatc gcaacacctt cctaaaatcg tcatatccca 36060 gataccatcc tggtataagc agcactatgt ctgatttccc ggcgaaggca ttgtgcattt 36120 tatggtatac accatactcg tatctcgcat cgtggaactt cggtacacca ccatgcgatc 36180 cataccacgt gattctcgaa ggaagtctgt tcggatttcc ggcgcttttc ggagcttcca 36240 gtttagtagc cggaggcatt gccacacagg accgattgac caaatcatgg caaccttctg 36300 atttcgtacc cgaaaatgcg gaggccccga ggagcacccc gaataactga aacagaacgc 36360 aaaatttaga ccggtaccct cccatacgat tatcacgtcc caaagccaaa tatcacggta 36420 ctcaccagcg caaaaatctt catgaagatc tggggcaaga gccgcacaac agtccacctg 36480 aatctgagac agcttaccgg cgactgtacc aaaccggtgt gccaccgccg aatttattct 36540 ctcaatctat taatgtttaa ctaaacaaac ttgctgacac ttgataaata tttactgacc 36600 tagacaattc cgagaacttc cttattaccg tgactatgca gtatcaagat aaacccttcc 36660 aaagttcgac tgagtacacg gtgtacttcc tgtcataaaa tatcacttct aaatatagtc 36720 ctaggtagaa actataaatg gtaagagaca acacactgat tcttagaaaa acctcacaga 36780 acaagatgaa cctcacatta tcctgactct tccttacgaa ctgaaataac cggtaagtca 36840 taaaaatgat tttccttttg acattgcgta cgcggaagca aactagaaaa atcgaacttg 36900 gaattttcca agtcgagtta actattaact tgatgacgtc aattggatta atcattaact 36960 cgaattagtt aatgattaac tagatctggt taatgattaa ctagtgacgt cactagttaa 37020 tcattaagct ttctatgcat atgcatgaga gattcatgca tatgaatgag agtctcatcc 37080 atattcataa acctcatcca tattcatgag cctctcatgc atatgcatgt ttttacatgc 37140 atatgcatgg acctcattca tattcatcaa ttcgttaatg tataacgcta tgactcactg 37200 tatatgttta cgttgcctag caacgttaat gatttacctg ctgacgtggc agctccgcct 37260 cccggtaaat catttacctg gactttgttc tttatgttta ttcaccatgg caacgctacc 37320 atatatggac atccgactcc gcctccccag ttatacatta acgatggcgt gataggcgga 37380 gctctctccc attggctctt aatgacgtag tccgggttaa ccataagcca gaaccgccta 37440 tataggtaga gcaggtagac ccggaacacc attcccatac ggacctccat agagtgcgga 37500 cctctacggg ctctccatac cggtaaatat tttattccat ttaatccaat caaataaatc 37560 aataatcaac tcaatgctgt gattctgcct caaattcaat ggtgattttc tttaataaaa 37620 agcccacccc ccttggcacc cccctgtaca cccccctgta caggcgacca ccccctatgg 37680 acacccccct gtacaggcga ccacccccta tggacacccc cctgtacagg cgaccacccc 37740 ctatggacac ccccctgtac aggcgaccac cccctatgga cacccccctg tacaggcgac 37800 caccccctat ggacaccccc ctgtacaccc ccctgtacat tttctcccat aggctacaat 37860 ggaacactgc cccctagtgt ctcccgctcc atgggacccc tatgaggtgg gcaccatctc 37920 atttgccgca atggagctca tccacgaggg ggcgccattg aagctagggg accgcataga 37980 gagcctggcc aatggggtgc tttggaatcc ggatatcccc gcccaactct tcaactgcat 38040 ttctattcgc tcatggggat cacatgggaa gcgcgtcata ttcaaaggcc acacccaccg 38100 gatgttccac gcccagctta ggatccgtag caccgccccc gttactagga aacaggccgg 38160 aagcctactc ctcagcctat cacagaggct cctgtgtttc cccgcccgct ataataccca 38220 ccccctcgtg atgcaattgg gggtggagtc taatgccatg ggacttccta tatattccaa 38280 gagggccctc gagatggcgc tcaagagcat gcgggtgcgc attgcgccta acagccacca 38340 ggtggcgcca cctgaaatag gaaagacctg tacggtgaag cccctcaaag aagtggagac 38400 cctccagcag ggggtcttca gtaccaccga tctaaaaaga gcacttccag aatggacttt 38460 tcgccgactt tttaacgaaa ttccctatat ttgtggctgg aagattggca ctgctccaaa 38520 gggcggagaa atggatggtt acgctccacc ccgaatccag agccccgccc accgaaggac 38580 ggaaggaccc gcccactctc caagacctcg cccggctggg cgtggtcgaa aactgcctca 38640 agatgaggag gcggggcata agccctaggc accaccccta ctttcaataa accaatcaga 38700 acagagcact tgactcctcc tatttgtggg cggggctatg ggtacggtat agagcgcccc 38760 ctggcggccg ttggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 38820 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 38880 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 38940 tggcgcgggc cggtcaatca ttaacccggc gcccagcacg ggcgccccct gccggccgga 39000 gacagacact gcgtgtctgc ggagctcccc ctggcggccg agggccgcca ctgcacccgc 39060 gccggactta gtctctggcg cgggccggtc aatcattaac ccggcgccca gcacgggcgc 39120 cccctgccgg ccggagacag acactgcgtg tctgcggagc tccccctggc ggccgagggc 39180 cgccactgca cccgcgccgg acttagtctc tggcgcgggc cggtcaatca ttaacccggc 39240 gcccagcacg ggcgccccct gccggccgga gacagacact gcgtgtctgc ggagctcccc 39300 ctggcggccg agggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 39360 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 39420 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 39480 tggcgcgggc cggtcaatca ttaacccggc gcccagcacg ggcgccccct gccggccgga 39540 gacagacact gcgtgtctgc ggagctcccc ctggcggccg agggccgcca ctgcacccgc 39600 gccggactta gtctctggcg cgggccggtc aatcattaac ccggcgccca gcacgggcgc 39660 cccctgccgg ccggagacag acactgcgtg tctgcggagc tccccctggc ggccgagggc 39720 cgccactgca cccgcgccgg acttagtctc tggcgcgggc cggtcaatca ttaacccggc 39780 gcccagcacg ggcgccccct gccggccgga gacagacact gcgtgtctgc ggagctcccc 39840 ctggcggccg agggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 39900 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 39960 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 40020 tggcgcgggc cggtcaatca ttaacccggc gcccagcacg ggcgccccct gccggccgga 40080 gacagacact gcgtgtctgc ggagctcccc ctggcggccg agggccgcca ctgcacccgc 40140 gccggactta gtctctggcg cgggccggtc aatcattaac ccggcgccca gcacgggcgc 40200 cccctgccgg ccggagacag acactgcgtg tctgcggagc tccccctggc ggccgagggc 40260 cgccactgca cccgcgccgg acttagtctc tggcgcgggc cggtcaatca ttaacccggc 40320 gcccagcacg ggcgccccct gccggccgga gacagacact gcgtgtctgc ggagctcccc 40380 ctggcggccg agggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 40440 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 40500 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 40560 tgacaccgca tttactttca acaccttttt ttatttcagc acaccgaaat ctcgtccgtg 40620 taaaccttgt ccccaaacct cagtcccgta catcgaaccc atccacatat atctcgcaca 40680 gctccagtgc ttacaagata tttccctact ggataattag gatcttgaca ccacttattc 40740 aaatacatcc ggaaccactg tccgtctgga acttggcttc tgtagaactc ctcacgaaag 40800 tagcgcttca ttgctctaga acacttatac aaatgtttat ctctcaggtc gaacatcgtg 40860 aggcaaatca acagcgtcaa atgacaagca caagaaccgc ccggctgaag cccgacgcag 40920 taacccatag gatgtccgac accgatgtta gcttcttcgt ggtacggata ccacagctcc 40980 ttgatgtctt gaactagctt ccagaagatg gcatcgtgca tacaccacgg aggtacggca 41040 acaccaaaca ccacgtacaa aggcatctcc cgacactctg caattacaac tagcatcaga 41100 aatacatcca gccgctattt gaaaaaaaca acaggatctc acccaaaaag aaaaagtact 41160 taccccgaaa ccaccaggaa atccccctta cgtgataatc cggatctcgc tgacgctcaa 41220 agaagccgcc ttaaataccg ctccttatct ctcttcgatt acctcagtta cgcccatttc 41280 ttatcaggta agatacgcag gtaagatacg cccttccaca tcagcacgga atgtgctgac 41340 cagcttaccg cgcccgtttt taatatccgt taattagtaa cttaggtgaa tcaccaaacc 41400 acctgccggc atatatctac acccttgata cggcggcatc cattgaagtc gagcgccacc 41460 atggagtcaa ctaccgtatc ctcgattctg ctcctatcct tcttcgtatc ttccatcgaa 41520 tcctatccgc tcctgcataa cttcacggcg ctcacgggat ccgtgcttac tcttccctac 41580 aaaggacatg accgaccctt taaattcgaa tggcggctga cagatcagac caaagtagca 41640 atctcggatc ctagcaccgg catcaactac ccgtcgggtc cactcaaagg aagagtacat 41700 ctaaacggca gcgccctcat cgttacatcc ctacgactta gtgacgccgg tacctataca 41760 atcctctcag aggacaacac aggaactgaa atcggatact cctattacgt cgaagtcaga 41820 ggtatagaac tttcccctcc tttttctaca tgatatcccg ttccctcttt tttcttaccc 41880 ctagcttatt cttcctagaa ccgatgcaag ctcccaccct ctatacggat cgctacaatg 41940 actcatctgc catacgcctc acatgccaag cttccaacaa tctccaccgt aatatcacat 42000 accactggaa aacagattta atacagccaa ctaactcgac tcgatctatc accttaaaca 42060 tcgaagacta tatatccgtt acctgcacag tcactgatgg agtcagcaaa aacagcatca 42120 cgattatggt tccgttgaga ggtagtattc ggtttcccgt tatctcccta tactacgtaa 42180 tcatgcccac ccttttaacc cgtatcttta caacagatcc ttctccacct acgccttacg 42240 gtttgcatcc cactatcatt ttcctgagtg tcctctgcat gatcctcatc accgcaatta 42300 ccgtctactt tgtgaagaag cattgctgcg ataagcaata taaaataact tgcattaacc 42360 cttaccgaga atgcttcggt ggcgccggtc tcgtttaatc tcgtttaaat aaaaaccact 42420 ataatccccg cttcctataa tctctacttc ctatgattaa atgggtaatt aaatgatggg 42480 taattaagtg gtgtcaatga tcaaataatg attcggaaac gatttcggga aaaatgattt 42540 taggaatcaa tgatcattat caaaaataaa tgatttattt ataagacgtg tttttgatga 42600 ttgatacgat tagtaaatca catgattaga aacacatgat tagaaacaca gtttaatcat 42660 caatcgaaat caccgcgcaa gatatggaca gtcacgttct cctctgatgt cctatccata 42720 gattcgctgt ccgtagaatc agaggcgcag agcatccgcc gaaaccagaa agttatcgct 42780 accatcagca ataacaacat aattaacgcg atcggcagcc acactggaaa ctcgccggtc 42840 tgattggata ccagtcctcc gccggtctta taagtggtct cctcgatctt ctgatcctcc 42900 tgagtcaaac caggaattac cactacgtaa aatccggtct ttgaatcctg ttgtaagtct 42960 agcgtattag tatacagtcc cgaatcggca ggtgttacgt ctctaatagt aacgcaattg 43020 gatgtggcat tgtaatgcgt gcggctctta tagttagcat acactatggg gttacccaga 43080 gctttataaa gctgtaatac gaacacggta gacgcgtgct cgttagagga ctgaaactcc 43140 cactccgtta tagaagcgga gctagcatcc ttcccacaca tgcgtaactc tccaccttcc 43200 aaaactaata tacggctcat gttatgcacc tcaaccgtcg cattgcccgt aatatccttc 43260 acggtatact gtatggcata cggtttccca ccgacaaagg aaccgtaagt caaccagcac 43320 ccgttattag ccttcagctt actcaggtaa atagtattgc aaccactatc taccctcgtt 43380 ctacccatgt acccgctttc aagtccaggt tctttaaccc tcggaatcac aaatgccgcc 43440 ctctccatct tattgcaatt gggggacggg gaatgatacc agaggatcat agtatggtcc 43500 ctgcactctc ctgtccgtag agccagctcc gggtcccggg gagcgccaaa gcagagtccg 43560 aggacagcag ccagcagcag ggcaacggac ttcatgatga atgcgggaac cagccagcag 43620 cagcaggtag aaggtgcgag tactcgaggc aggcgggaat aacaccgcac gctttccacc 43680 ccgcttaaat acacaaccca caccggtcat ggtcaaacat tacctagcct actcagcacg 43740 gaaaagtact cacccttaac acagttcgac ttatcaggta cacacccgta accttcgcac 43800 cccaaagcgc acagtcagca ccataaggaa gtcgtaaagt taccctttga ttgcattgtc 43860 atcataaaaa acgctgagca aacggaaacg ccccaagtac aaatctctct tccgccacgc 43920 cctcgttaat caataactag tcgccgatac atatataccc cctccgttcc atcacaccta 43980 cactaccttc tctccgcaca ggcaacatct caatccttac tcttccgaat ccacttcgcc 44040 gccttcgaca tgaatccggt aagtatcaac ttttttctaa gcttatgctt ctgacacctc 44100 tgatcttaat accgcacccc ttttttctca aaaaaggttt cgtgcttaac gcttcttcta 44160 tgcgtcatca tcccgagagc cgatccgctt ccaatcaccc ccgcctctaa acccgctacc 44220 gactccgccc ggaccggcgc ctccgtcatc acgacccact tacccgcatc tccgagcgcc 44280 accccgtgcg acgaaatctt gagcgaggac tgttggttcg aaaatgccag cggtgattac 44340 caacccctac cctgggaacc gaaagaggaa acggtttcag atcaaaaccc tctcaccgca 44400 accgacccca tcggtgaccg aatccccgct atcatccagt cccagtcccg cgcttctaac 44460 cgtccgacca gtaaggaaca cacttctact atcgcatcca tctcgaccgt agtcggcatc 44520 gcggtcctcg tcatcctgac cctcgtagcc tactttacca agtaccccaa accgagaccg 44580 cccagatcca tctacatagg agtagctcca cccgatatgg aactcaagga aatataattc 44640 cagccccacc cttaacccta cctttccatg agtcagtatt ttcaataaag ttatattgca 44700 gtattaattc cgttgtttcc gcgttctttc tctcgcgcgg acaaagtccg ttcgaccagg 44760 aagtccggta tacgtcattc cgcggtgtca tgatgacgcg cataactcac gactgccatc 44820 tgccggacaa acgcggtact acacttaaca cataaacacc cgcctttttt cgattcccac 44880 cataatcagg ctttggtaaa aattcgcaga ttctaaaatc cgtattcctg cgcccgcgat 44940 aatagatcac gcccacgaca cgccctagcg ctcttatcac acggtctctg ctgccatcta 45000 gcgggcggga gcggtagtgc gaggtgacag cagcgtcatt catgtaggta tatatagatg 45060 atg 45063 <210> SEQ ID NO 2 <211> LENGTH: 714 <212> TYPE: DNA <213> ORGANISM: Aequorea victoria <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(714) <223> OTHER INFORMATION: Enhanced green fluorescent protein (EGFP) <400> SEQUENCE: 2 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 60 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 120 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 180 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 240 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 300 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 360 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 420 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 480 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 540 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 600 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 660 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caag 714 <210> SEQ ID NO 3 <211> LENGTH: 711 <212> TYPE: DNA <213> ORGANISM: Discosoma sp <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(711) <223> OTHER INFORMATION: mCherry Red protein <400> SEQUENCE: 3 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag 60 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc 120 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc 180 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac 240 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc 300 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac 360 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta 420 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc 480 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct 540 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc 600 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa 660 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta g 711 <210> SEQ ID NO 4 <211> LENGTH: 553 <212> TYPE: DNA <213> ORGANISM: Human cytomegalovirus <220> FEATURE: <221> NAME/KEY: promoter <222> LOCATION: (1)..(553) <223> OTHER INFORMATION: Immeadiate early promoter <400> SEQUENCE: 4 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 60 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 120 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 180 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 240 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 300 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 360 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 420 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 480 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 540 ggtctatata agc 553 <210> SEQ ID NO 5 <211> LENGTH: 584 <212> TYPE: DNA <213> ORGANISM: Cytomegalovirus <220> FEATURE: <221> NAME/KEY: enhancer <222> LOCATION: (1)..(584) <223> OTHER INFORMATION: CMV enhancer/ chicken beta-actin promoter (CAG) <400> SEQUENCE: 5 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 60 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 120 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 180 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 240 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 300 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 360 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 420 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 480 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 540 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcg 584 <210> SEQ ID NO 6 <211> LENGTH: 1190 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: promoter <222> LOCATION: (1)..(1190) <223> OTHER INFORMATION: Human elongation factor 1 alpha (EF1A) promoter <400> SEQUENCE: 6 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 60 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg 120 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 180 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa 240 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt 300 gaattacttc cacgcccctg gctgcagtac gtgattcttg atcccgagct tcgggttgga 360 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt 420 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt 480 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt 540 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt 600 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg 660 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct 720 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaac aggctggccc 780 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct 840 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa 900 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca 960 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt 1020 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc 1080 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca 1140 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga 1190 <210> SEQ ID NO 7 <211> LENGTH: 114 <212> TYPE: DNA <213> ORGANISM: Fowlpox virus <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(114) <223> OTHER INFORMATION: Promoter Sequence for L2R (Fowlpox promoter) <400> SEQUENCE: 7 catagtaaat atgggtaact tcttaatagc cataattaaa attgaaaaaa aaatatcatt 60 ataaaacgta aacgaacaaa aaacattaat ttaaatttgc caataacaaa ccat 114 <210> SEQ ID NO 8 <211> LENGTH: 276 <212> TYPE: DNA <213> ORGANISM: eukaryotic cells <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(276) <223> OTHER INFORMATION: Promoter Sequence for B-actin (minimal) <400> SEQUENCE: 8 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa 60 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg 120 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg 180 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg 240 cggcggccct ataaaaagcg aagcgcgcgg cgggcg 276 <210> SEQ ID NO 9 <211> LENGTH: 516 <212> TYPE: DNA <213> ORGANISM: Woodchuck hepatitis virus <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(516) <223> OTHER INFORMATION: Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) <400> SEQUENCE: 9 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 60 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 120 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 180 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 240 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 300 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 360 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc 420 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 480 tccagcggac cttccttccc gcggcctgct gccggc 516 <210> SEQ ID NO 10 <211> LENGTH: 1695 <212> TYPE: DNA <213> ORGANISM: Influenza A virus (A/chicken/Mexico/435/2005 (H5N2)) <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1695) <223> OTHER INFORMATION: Segment 4 Hemagglutinin gene <400> SEQUENCE: 10 atggaaagaa tagtgattgc ctttgcaata atcagcattg ttacaggtga ccaaatctgc 60 attggttatc atgcaaacaa ttcaacaaaa caagttgata caatcatgga aaagaatgtg 120 acggtcacac atgctcagga catactagaa aaagaacaca atggaagact ctgcagtctt 180 aaaggagtaa agcccctcat cctgaaggat tgcagcgtag ctgggtggat tctgggaaat 240 cctatgtgtg gtgaattcct gaatgtgccg gaatggtcat acattgtgga gaaagacaac 300 ccatccaatg gcctgtgtta ccctggaaac ttcgacaatt atgaagaatt gaagcattta 360 atgagcagca caaatcacat cgagagaatt cagatatttc ctaggagctc ttggtccaac 420 cataatgcct catcaggagt gagctcggca tgcccgtcca atggtatatc ttcctttttc 480 cggaatgtag tgtggttgat caagaaggat aatgtgtacc gaacactaaa gaagaactac 540 accaacacca atgtagaaga ccttttaata atgtggggag ttcatcaccc tactgatgca 600 actgagcaga caaaactcta ccagaacctg aacacttatg tatcggtggg aacatcaaca 660 ctaaatcaga ggtcaatccc gaaaatagcc accagacccc aggtgaacgg acagagagga 720 aggatggaat ttttttggac aatattaaag ccgaacgatt caatcttttt tgagagtact 780 gggaacttta tagctcccga atatgcatac aagatcacta aaaaagggaa atcagcagtc 840 atgaaaagtg aactggatta tggcgactgt aataccaaat gccagacccc agtgggtgct 900 gtaaattcca gtctgccttt ccacaatgtc catcccttta ccattgggga atgccccagg 960 tatgtaaaat cgaagaaact ggtccttgca acaggattaa gaaatgtacc ccaaaaagaa 1020 acaagaggct tatttggagc aatagctgga ttcatagaag gggggtggca aggaatggtg 1080 gatggatggt atggatacca tcatagcaat gagcagggta gtgggtatgc tgcagacaaa 1140 gaatctacac agaaagcaat taatggaatc actaataaag tcaactcaat cattggcaaa 1200 atgaacactc agttcgaagc cattgggaaa gaattcaaca acctagaaag gagaatagaa 1260 aatttgaata agaaaatgga agatgacttt ttggatgtat ggacttacaa tgcagaactt 1320 ctagtgctca tggaaaacga gagaactctg gatctccatg attcaaatgt caagaattta 1380 tacgataagg tccgactcca actgagagat aatgcaaaag aactaggcaa cggatgcttt 1440 gaattctacc acaagtgtga caatgaatgc atggaaagtg tgagaaatgg aacgtatgat 1500 tacccacaat actcaaaaga gtcaaaactg aatagagagg aaatagaagg ggtcaaatta 1560 gaatcaacag ggacttatca gatactttca atctattcaa cagtggcgag ttccctagca 1620 ctggcaatca tggtagctgg tctatctttt tggatgtgct ccaatggatc attgcagtgc 1680 agaatttgca tctaa 1695 <210> SEQ ID NO 11 <211> LENGTH: 1723 <212> TYPE: DNA <213> ORGANISM: Influenza A virus (A/chicken/Jalisco/CPA1/2012(H7N3)) <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1722) <223> OTHER INFORMATION: Segment 4 Hemagglutinin gene <400> SEQUENCE: 11 ggatacaaaa tgaacactca aattttggca ctcattgctt gtatgctgat tggagctaaa 60 ggagataaaa tatgtcttgg gcaccatgct gtggcaaatg gaacaaaagt gaacacatta 120 acagagagag gaatcgaagt agtaaatgcc acagaaacgg tggagactgc aaatactaag 180 aaaatatgca ctcaggggaa aagaccaaca gatctgggac aatgcggact tctaggaacc 240 ctaataggac ctccccaatg cgatcaattt ctggaatttg acgctgattt aataattgaa 300 cgaagagaag gaaccgatgt gtgttatccc gggaagttca caaatgaaga atcactgagg 360 caaatccttc gagggtcagg aggaattgat aaagagtcaa tgggtttcac ctatagtgga 420 ataagaacca atggggcgac aagtgcttgc agaagatcag gttcttcctt ctatgcggag 480 atgaagtggt tactgtcgaa ttcagacaat gcggcttttc cccaaatgac taagtcgtac 540 agaaatccca ggaacaaacc agctctgata atttggggag tgcatcattc tggatcggct 600 actgagcaga ccaaactcta tgggagtgga aacaagttga taacagtagg aagctcgaaa 660 taccagcagt cattcacccc aagcccgggg gcacgaccac aggtgaatgg gcaatcagga 720 aggattgatt ttcactggct actccttgac cccaatgaca cagtgacctt cactttcaat 780 ggggcattca tagcccctga cagagcaagt ttctttagag gagagtcaat aggagttcag 840 agtgatgttc ctttggattc tggttgtgag ggggattgct tccacaatgg gggtacgata 900 gtgagttccc tgccattcca gaacatcaac cctagaacag tgggaaaatg ccctcgatat 960 gtcaaacaga caagcctcct tttggctaca gggatgagaa acgtcccaga gaaccccaag 1020 gataggaaga gccgacatcg aaggaccaga ggcctttttg gagcgattgc tggattcata 1080 gagaatggat gggaaggtct cattgatgga tggtatggtt tcagacatca aaatgcacaa 1140 ggagaaggaa ctgcagctga ttacaaaagc actcaatctg caatagatca gatcacaggc 1200 aaattgaatc gtctaattga caaaacaaat cagcagtttg aactgataga caacgaattc 1260 agtgaaatag aacaacaaat tgggaatgtc attaactgga cacgagattc aatgactgag 1320 gtatggtcgt acaatgctga attgctggta gctatggaaa atcagcacac aatagatctt 1380 gcagactcag aaatgaacaa actttatgag cgtgtaagga aacaactgag ggagaatgct 1440 gaagaggatg ggactggatg ctttgagata tttcataagt gtgatgatca gtgcatggag 1500 agcatcagga acaacactta tgaccatact caatacagag cggagtcatt gcagaataga 1560 atacagatag acccagtgaa attgagtagt ggatacaaag acataatctt atggtttagc 1620 ttcggggcat catgttttct tcttctagcc attgcaatgg gattggtttt catttgcata 1680 aagaatggaa acatgcggtg cactatttgt atatagtttg aga 1723 <210> SEQ ID NO 12 <211> LENGTH: 1723 <212> TYPE: DNA <213> ORGANISM: Influenza A virus (A/chicken/Guanajuato/07437-15/ 2015(H7N3)) <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1722) <223> OTHER INFORMATION: Segment 4 Hemagglutinin gene <400> SEQUENCE: 12 ggatacaaaa tgaacactca aattttggca ctcattgctt atatgctaat tggagctaaa 60 ggagacaaaa tatgtcttgg gcaccatgct gtggcaaatg gaacaaaagt gaacacatta 120 acagagaggg gaatcgaagt agtaaatgcc acagaaacgg tggagactgc aaatactaag 180 aaaatatgca ctcaggggaa aagaccaata gatctaagac aatgcggact tctaggaacc 240 ctaataggac ctccccaatg cgatcaattt ctgaaatttg acgctgattt aataattgaa 300 cgaagagaag gaaccgatgt gtgttatccc gggaagttca caaatgaaga atcactgagg 360 caaatacttc gaagatcagg aggaattgat aaagagtcga tggatttcac ctataatgga 420 ataagaacca atgggacgac aagtgcttgc aaaagatcca atccttcctt ctatgcggag 480 atgaagtggt tactgtcgga ttctgacaat gcaactttcc ctcaaatgac tatgtcgtac 540 agaaatccca ggaacaaatc agctctgata atttggggag tgcatcattc tggatcggct 600 actgagcaga ccaaactcta tgggagtgga gacaagttca taacagtagg aagctcgaaa 660 tacctgcagt cattcacccc aagcccgggg ccacgaccaa aggtgaatgg gcaatcagga 720 aggatcgact ttcactggct actccttgat cccaatgaca cagtgacctt cactttcaat 780 ggggcattca tagctcctga cagagcaagc ttctttagag gagagtcaat aggaattcag 840 agtgatgttc ctttggatcc tggttgtaag ggggattgct tccacaatgg gggtacgata 900 gtgagttctc tgccattcca gaacatcaac cctagaacag tgggaaaatg ccctcggtat 960 gtcaaacagc caagcctcct tttggctaca gggatgagaa atgtcccaga gaaccccaag 1020 gataggaaga gccgacatcg aaggaccaga ggcctttttg gagcgattgc tgggttcata 1080 gagaatggat gggaaggtct cattgatgga tggtatggtt tcagacatca aaatgcacaa 1140 ggagaaggaa ctgcagctga ttacaaaagc actcaatctg cgataaatca aatcacaggc 1200 aaattgaatc gtctaattga caaaacaaat cagcagtttg aactgataga caacgaattc 1260 agtgaaatag aacaacagat tgggaatgtc attaactgga cacgcgattc aatgactgag 1320 gtatggtcgt acaatgctga attgctgata gctatggaaa atcagcacac aatagatctt 1380 gcagactcag aaatgaacaa actttatgag cgtgtaagga aacaacttag ggagaatgct 1440 gaagaggatg ggactggatg ctttgaaatg tttcataagt gtgatgatca gtgcatggag 1500 agcatcagga acaacactta tgaccatact caatacagag cggaatcatt gcagaatagg 1560 atacagatag atccagtgaa attgagtagt ggatacaaag acataatctt atggtttagc 1620 ttcggggcat catgttttct tcttctagcc attgcaatgg gattggtttt catttgcata 1680 aagaatggaa acatgcggtg cactatttgt atatagtttg aga 1723 <210> SEQ ID NO 13 <211> LENGTH: 2652 <212> TYPE: DNA <213> ORGANISM: Gallid herpesvirus 1 strain USDA <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(2652) <223> OTHER INFORMATION: Laryngotracheitis virus (ILT) glycoprotein B <400> SEQUENCE: 13 atgcaatcct acatcgccgt gaacattgac atggctagct tgaaaatgct gatctgcgtg 60 tgcgtggcaa tcctgatccc atctacccta tctcaagatt cacacggaat tgctggaata 120 atagaccctc gtgatacagc cagcatggat gttggaaaaa tctctttctc cgaagccatt 180 gggtcggggg caccgaaaga accccagatt agaaacagaa tttttgcgtg ctcatctcca 240 actggcgcca gtgttgcgag gcttgcccag ccacgacatt gtcaccgaca tgccgattcg 300 actaacatga ctgaaggaat tgccgtagtc ttcaagcaaa acattgcccc gtacgtcttt 360 aatgtgactc tatactataa acatataacc acagttacta cgtgggcatt attctcaaga 420 ccccaaataa caaatgagta cgtgaccagg gttccaatag actatcatga aattgtcagg 480 attgatcgat cgggagaatg ctcatccaaa gcaacgtatc ataaaaattt catgtttttt 540 gaagcttacg acaatgatga agcagaaaaa aaattgcccc tggttccatc actgttaaga 600 tcaactgtct ccaaggcgtt tcatacaact aactttacta agcgacatca aaccctggga 660 taccgaacgt ctacatcggt cgactgtgtt gtgaaatatc tacaggctag atctgtatac 720 ccgtatgatt actttggaat ggcgacaggt gatacagtag aaatttctcc tttttatacc 780 aaaaacacga ccggaccaag gcgtcacagt gtctacagag actatagatt tctcgaaatc 840 gcaaattatc aagtcaggga tttggaaacc ggacaaataa gaccccctaa aaaaagaaac 900 tttctaacag atgaacaatt cactataggc tgggatgcaa tggaagaaaa ggaatctgta 960 tgtactctca gtaaatggat tgaagtcccg gaagcagttc gtgtttcgta caaaaacagt 1020 taccactttt cacttaaaga tatgactatg acgttctcgt ccggaaaaca accttttaac 1080 atcagcaggc ttcatttggc tgaatgcgtt cctaccatag cctcggaggc catagatggc 1140 atctttgcca gaaagtatag ttcgactcat gtccgttctg gggacatcga atactatctc 1200 ggtagtggcg gatttctgat cgcatttcag aaactcatga gccatggctt ggctgaaatg 1260 tacctagaag aggcacaaag acaaaatcat ctcccgagag ggagagagcg tcgccaagcc 1320 gcaggtcgcc gcacggcgtc gctgcagtct ggacctcagg gtgatagaat tactacccac 1380 agttctgcaa catttgccat gttacaattt gcatacgaca aaatccaagc ccatgttaac 1440 gagcttatcg gaaatttgtt ggaagcgtgg tgtgagcttc agaaccgcca actgattgta 1500 tggcatgaga tgaagaaact aaacccgaac tcactgatga catctttgtt cggacaacct 1560 gtaagcgcca ggctattggg agacatcgta gcggtatcaa aatgtataga aattccaatc 1620 gaaaatatta ggatgcagga ttccatgcgc atgccagggg acccaaccat gtgctatacc 1680 agaccagtac ttattttcag gtattcgtcc tcccctgagt cacagttttc tgcgaactca 1740 acagaaaacc acaatcttga catattaggc caactcggag aacataatga aattttacaa 1800 gggcggaatt tgatagaacc atgcatgatc aatcacagac ggtactttct gttgggagaa 1860 aactaccttc tttacgaaga ctatacattt gttagacaag taaatgcttc cgagatcgaa 1920 gaagtgagca cattcatcaa cttgaacgcc actatactag aagatttgga ctttgtgccc 1980 gtcgaagtat acactcgcga ggaactcaga gatactggga ctttaaacta tgatgatgtg 2040 gtcagatatc aaaatattta taacaaaagg ttcagagaca ttgacactgt aatacgtgga 2100 gataggggag atgcaatctt tagagcaata gcagattttt ttggcaacac tcttggagaa 2160 gtaggaaagg cattgggaac tgtagtgatg acagccgcgg cagcagtaat ttctacagta 2220 tctggcatcg cctcatttct ttctaacccg ttcgccgcac tcggaattgg gatagcggtg 2280 gtggtgagca ttattttagg actgctggcg ttcaaatatg taatgaacct gaaatcaaac 2340 ccagttcagg ttctgttccc aggcgcagtt cccccggccg gaactcctcc acgaccctct 2400 agacgttact acaaggatga ggaggaggtt gaggaggata gtgatgagga cgacaggata 2460 cttgccacca gagttctgaa aggccttgag cttctacaca aggatgaaca gaaagctcga 2520 agacagaaag cgcggttttc tgcttttgct aaaaatatga gaaacctatt tcgcagaaaa 2580 ccccgaacca aggaagatga ctaccccctg ctcgaatacc cttcgtgggc agaagaaagc 2640 gaagacgaat aa 2652 <210> SEQ ID NO 14 <211> LENGTH: 1305 <212> TYPE: DNA <213> ORGANISM: Gallid herpesvirus 1 strain USDA <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1305) <223> OTHER INFORMATION: Laryngotracheitis virus (ILT) glycoprotein D <400> SEQUENCE: 14 atgcaccgtc ctcatctcag acggcactcg cgttactacg cgaaaggaga ggtgcttaac 60 aaacacatgg attgcggtgg aaaacggtgc tgctcaggcg cagctgtatt cactcttttc 120 tggacttgtg tcaggattat gcgggagcat atctgctttg tacgcaacgc tatggaccgc 180 catttatttt tgaggaatgc tttttggact atcgtactgc tttcttcctt cgctagccag 240 agcaccgccg ccgtcacgta cgactacatt ttaggccgtc gcgcgctcga cgcgctaacc 300 ataccggcgg ttggcccgta taacagatac ctcactaggg tatcaagagg ctgcgacgtt 360 gtcgagctca acccgatttc taacgtggac gacatgatat cggcggccaa agaaaaagag 420 aaggggggcc ctttcgaggc ctccgtcgtc tggttctacg tgattaaggg cgacgacggc 480 gaggacaagt actgtccaat ctatagaaaa gagtacaggg aatgtggcga cgtacaactg 540 ctatctgaat gcgccgttca atctgcacag atgtgggcag tggactatgt tcctagcacc 600 cttgtatcgc gaaatggcgc gggactgact atattctccc ccactgctgc gctctctggc 660 caatacttgc tgaccctgaa aatcgggaga tttgcgcaaa cagctctcgt aactctagaa 720 gttaacgatc gctgtttaaa gatcgggtcg cagcttaact ttttaccgtc gaaatgctgg 780 acaacagaac agtatcagac tggatttcaa ggcgaacacc tttatccgat cgcagacacc 840 aatacacgac acgcggacga cgtatatcgg ggatacgaag atattctgca gcgctggaat 900 aatttgctga ggaaaaagaa tcctagcgcg ccagaccctc gtccagatag cgtcccgcaa 960 gaaattcccg ctgtaaccaa gaaagcggaa gggcgcaccc cggacgcaga aagcagcgaa 1020 aagaaggccc ctccagaaga ctcggaggac gacatgcagg cagaggcttc tggagaaaat 1080 cctgccgccc tccccgaaga cgacgaagtc cccgaggaca ccgagcacga tgatccaaac 1140 tcggatcctg actattacaa tgacatgccc gccgtgatcc cggtggagga gactactaaa 1200 agttctaatg ccgtctccat gcccatattc gcggcgttcg tagcctgcgc ggtcgcgctc 1260 gtggggctac tggtttggag catcgtaaaa tgcgcgcgta gctaa 1305 <210> SEQ ID NO 15 <211> LENGTH: 1734 <212> TYPE: DNA <213> ORGANISM: Newcastle disease virus strain La Sota C5 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1734) <223> OTHER INFORMATION: HN gene <400> SEQUENCE: 15 atggaccgcg ccgttagcca agttgcgtta gagaatgatg aaagagaggc aaaaaataca 60 tggcgcttga tattccggat tgcaatctta ttcttaacag tagtgacctt ggctatatct 120 gtagcctccc ttttatatag catgggggct agcacaccta gcgatcttgt aggcataccg 180 actaggattt ccagggcaga agaaaagatt acatctacac ttggttccaa tcaagatgta 240 gtagatagga tatataagca agtggccctt gagtctccgt tggcattgtt aaatactgag 300 accacaatta tgaacgcaat aacatctctc tcttatcaga ttaatggagc tgcaaacaac 360 agtgggtggg gggcacctat ccatgaccca gattatatag gggggatagg caaagaactc 420 attgtagatg atgctagtga tgtcacatca ttctatccct ctgcatttca agaacatctg 480 aattttatcc cggcgcctac tacaggatca ggttgcactc gaataccctc atttgacatg 540 agtgctaccc attactgcta cacccataat gtaatattgt ctggatgcag agatcactca 600 cattcatatc agtatttagc acttggtgtg ctccggacat ctgcaacagg gagggtattc 660 ttttctactc tgcgttccat caacctggac gacacccaaa atcggaagtc ttgcagtgtg 720 agtgcaactc ccctgggttg tgatatgctg tgctcgaaag tcacggagac agaggaagaa 780 gattataact cagctgtccc tacgcggatg gtacatggga ggttagggtt cgacggccag 840 taccacgaaa aggacctaga tgtcacaaca ttattcgggg actgggtggc caactaccca 900 ggagtagggg gtggatcttt tattgacagc cgcgtatggt tctcagtcta cggagggtta 960 aaacccaatt cacccagtga cactgtacag gaagggaaat atgtgatata caagcgatac 1020 aatgacacat gcccagatga gcaagactac cagattcgaa tggccaagtc ttcgtataag 1080 cctggacggt ttggtgggaa acgcatacag caggctatct tatctatcaa ggtgtcaaca 1140 tccttaggcg aagacccggt actgactgta ccgcccaaca cagtcacact catgggggcc 1200 gaaggcagaa ttctcacagt agggacatct catttcttgt atcaacgagg gtcatcatac 1260 ttctctcccg cgttattata tcctatgaca gtcagcaaca aaacagccac tcttcatagt 1320 ccttatacat tcaatgcctt cactcggcca ggtagtatcc cttgccaggc ttcagcaaga 1380 tgccccaact cgtgtgttac tggagtctat acagatccat atcccctaat cttctataga 1440 aaccacacct tgcgaggggt attcgggaca atgcttgatg gtgtacaagc aagacttaac 1500 cctgcgtctg cagtattcga tagcacatcc cgcagtcgca ttactcgagt gagttcaagc 1560 agtaccaaag cagcatacac aacatcaact tgttttaaag tggtcaagac taataagacc 1620 tattgtctca gcattgctga aatatctaat actctcttcg gagaattcag aatcgtcccg 1680 ttactagttg agatcctcaa agatgacggg gttagagaag ccaggtctgg ctag 1734 <210> SEQ ID NO 16 <211> LENGTH: 1716 <212> TYPE: DNA <213> ORGANISM: Newcastle disease virus strain M08-3313/Mexico <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1716) <223> OTHER INFORMATION: Hemagglutinin neuramidase of Newcastle disease virus strain M08-3313/Mexico <400> SEQUENCE: 16 atggatcgtg tagttagcag agttgtacta gagaacgaag aaagagaagc aaagaacaca 60 tggcgcttgg ttttccggat cgcagtctta tctctaatag taatgacttt agctatctct 120 gtagccgccc tggtatacag catgggggct agcacaccga gcgaccttgc aggcatatca 180 acggtgatct ctaaggcaga ggatagggtt acatctttac tcagttcaaa tcaagatgtg 240 gtagataggg tatataaaca ggtggccctt gagtccccgc tggcgttgct aaatactgaa 300 tctataatta tgaatgcaat aacttctctc tcttatcaaa ttaatggggc tgcaaataat 360 agtgggtgtg gggcacctgt tcatgaccca gattatattg ggggggtagg caaagaactc 420 atagtagatg acacaagtga tgtcacatca ttctatcctt cagcatacca agaacacctg 480 aattttatcc cggcgcctac tacaggatca ggctgcactc ggataccctc attcgacatg 540 agcgctaccc actattgtta tactcacaat gtgatattat ctggttgcag agatcactcg 600 cactcacatc agtatttagc gctaggtgta cttcggacat ctgcaacagg gagggtattc 660 ttttctactc tgcgttccat caatttagat gacacccaaa atcggaagtc ttgcagtgtg 720 agtgcgactc ctttaggttg tgatatgctg tgctctaaag tcatagagac tgaggaggag 780 gattataagt caattacccc cacatcaatg gtgcatggaa ggttagggtt tgacggtcag 840 caccatgaga aggacttaga cgtcacagtc ttatttaagg attgggttgc aaattacccg 900 ggagtgggag gagggtctct tattgacgac cgtgtatggt tcccagttta tggagggcta 960 aaacccaatt cgcctagcga cactgcacaa gaagggagat atgtaatata caagcgctat 1020 aataacacat gccccgatgg acaagattac caagttcgga tggctaagtc ttcgtataag 1080 cctggacggt ttggtagaaa gcgcgtacag caagccatct tatctatcaa agtatcaaca 1140 tccttgggcg aggacccggt gctgactgta ccgccaaata cagttacact catgggggct 1200 gaaggcagag tcctcacagt agggacatct catttcttgt accaacgagg gtcttcatac 1260 ttctctcccg ccttattata ccctatgaca gtacacaaca aaacagctac tcttcatagt 1320 ccttatatat ttaatgcttt cactcggcca ggtagtgtcc cttgccaggc atcagcaagg 1380 tgccctaact catgtatcac tggagtctat actgatccgt atcctgtagt cttccatagg 1440 aatcacacct tgcgaggggt gttcgggaca atgcttgata atgaacaagc aaggttcaac 1500 cccgtatctg cagtatttga ttacacatct cgcagtcgca taacccgggt aagttcaagc 1560 agcaccaagg cagcatacac gacatcgaca tgttttaaag ttgtcaagac caataaagtt 1620 tattgcctta gcattgcaga aatatccaac accctatttg gggaattcag gattgtccct 1680 ttactggtcg agatcctcaa ggatgatagg gtttaa 1716 <210> SEQ ID NO 17 <211> LENGTH: 1350 <212> TYPE: DNA <213> ORGANISM: Chicken anemia virus isolate CAV-10 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1350) <223> OTHER INFORMATION: VP1 gene of Chicken anemia virus isolate CAV-10 <400> SEQUENCE: 17 atggcaagac gagctcgcag accgagaggc agattttacg ccttcagaag aggacggtgg 60 caccacctca agcgacttcg acgaagatat aaatttcgac atcggaggag acagcggtat 120 cgtagacgag cttttaggaa ggcctttcac aacccccgcc ccggtacgta tagtgtgagg 180 ctaccgaacc cccagtcaac gatgactatc cgctttcaag gcatcatttt tcttaccgaa 240 ggactcattc tacctaaaaa cagtacagcg gggggatatg cagaccacct ctacggggcg 300 agagtcgcca agatctcagt gaacctgaaa gagttcctgc tagcgtcaat gaacctgaca 360 tacgtgagca aaataggagg ccccatcgcc ggtgagttga ttgcggacgg gtctcaatca 420 caagccgcgc aaaactggcc taattgctgg ctgccgctag ataataacgt gccctccgct 480 acaccatcgg catggtggag atgggcttta atgatgatgc agcccacgga ctcttgccgg 540 ttctttaatc accctaaaca aatgaccctg caagacatgg gtcggatgtt tgggggctgg 600 catctgttcc gacacattga aacccgcttt cagctccttg ccactaagaa tgagggatcc 660 ttcagccccg tggcgagtct tctctcccag ggagagtacc tcacgcgtcg ggacgatgtt 720 aagtacagca gcgatcacca gaaccggtgg cgaaaaggcg agcaaccgat gacggggggt 780 attgcttatg cgacggggaa aatgagaccc gacgagcaac agtaccctgc tatgcccccc 840 gaccccccga tcatcacaac taccacaccg caaggcacgc aagtccgctg catgaatagc 900 acgcaagctt ggtggtcgtg ggacacatat atgagctttg caacactcac agcgctcggt 960 gcacaatggt ctttccctcc agggcaacgt tcagtttcta gacgctcgtt caaccaccac 1020 aaagcgagag gggccgggga ccccaaggga cagagatggc acacgcttgt tccgctcggc 1080 acagagacca tcaccgacag ctacatgagt gcgccggcat cagagctgga cactaacttc 1140 tttacgcttt acgtagcgca aggcacaaat aaatcgcagc agtacaagtt cggcacagct 1200 acatacgcgc taaaggaacc ggtaatgaag agcgatgcat gggcggtggt acgcgtccag 1260 tcagtctggc aactgggtaa caggcaaagg ccatacccat gggacgtcaa ctgggccaac 1320 agcaccatgt actgggggtc gcagccctga 1350 <210> SEQ ID NO 18 <211> LENGTH: 651 <212> TYPE: DNA <213> ORGANISM: Chicken anemia virus isolate CAV-10 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(651) <223> OTHER INFORMATION: VP2 gene of chicken anemia virus isolate CAV-10 <400> SEQUENCE: 18 atgcacggga acggcggaca accggccgct gggggcagtg aatcggcgct tagccgagag 60 gggcaacctg ggcccagcgg agccgcgcag gggcaagtaa tttcaaatga acgctctcca 120 agaagatact ccacccggac catcaacggt gttcaggcca ccaacaagtt cacggccgtt 180 ggaaacccct cactgcagag agatccggat tggtatcgct ggaattacaa tcactctatc 240 gctgtgtggc tgcgcgaatg ctcgcgctcc cacgctaaga tctgcaactg cggacaattc 300 agaaagcact ggtttcaaga atgtgccgga cttgaggacc gatcaaccca agcctccctc 360 gaagaagcga tcctgcgacc cctccgagta cagggtaagc gagctaaaag aaagcttgat 420 taccactact cccagccgac cccgaaccgc aagaaggtgt ataagactgt aagatggcaa 480 gacgagctcg cagaccgaga ggcagatttt acgccttcag aagaggacgg tggcaccacc 540 tcaagcgact tcgacgaaga tataaatttc gacatcggag gagacagcgg tatcgtagac 600 gagcttttag gaaggccttt cacaaccccc gccccggtac gtatagtgtg a 651 <210> SEQ ID NO 19 <211> LENGTH: 1482 <212> TYPE: DNA <213> ORGANISM: Fowl adenovirus 4 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1482) <223> OTHER INFORMATION: Short fiber gene <400> SEQUENCE: 19 ggtcctatcc ctttttccta tcagggttac gtctactccc ccaacgggaa caatgctccg 60 ggcccctaaa agaagacatt ccgaaaacgg gaagcccgag accgaagcgg gaccttcccc 120 ggctccaatc aagcgcgcca aacgcatggt gagagcatcc cagcttgacc tggtttatcc 180 tttcgattac gtggccgacc ccgtcggagg gctcaacccg ccttttttgg gaggctcagg 240 acccctagtg gaccagggcg gacagcttac gctcaacgtc accgatccca tcatcatcaa 300 gaacagatcg gtggacttgg cccacgaccc cagtctcgat gtcaacgccc aaggtcaact 360 ggcggtggcc gttgaccccg aaggggccct ggacatcacc cccgatggac tggacgtcaa 420 ggtcgacccg tccggcggat tggattccac cgcgggtgga ctgggggtca gcgtggacga 480 caccttgctc gtggatcagg gagaactggg cgtacacctc aaccaacaag gacccatcac 540 tgccgatagc agtggtatcg acctcgagat caatcctaac atgttcacgg tcaacacctc 600 gaccggaagc ggagtgctgg aactcaacct aaaagcgcag ggaggcatcc aagccgacag 660 ttcgggagtg ggcgtttccg tggatgaaag cctacagatt gtcaacaaca ctctggaagt 720 gaaaccggat cccagcggac cgcttacggt ctccgccaat ggcctagggc tgaagtacga 780 cactaatacc ctagcggtga ccgcgggcgc tttaaccgtg gtcggagggg ggagcgtctc 840 cacacccaat cgctactttt gtctcgggaa gtcacagcct caacacctac aatgccacgc 900 cgtcaattcc agcgcgaacg ccttctcttg cgcctactac cttcaacagt ggaacataca 960 ggggctcctt gttacctccc tctacttgaa attggacagc gccaccatgg ggaatcgccc 1020 tggggacctc aactccgcca atgccaaatg gttcaccttt tgggtgtccg cctatctcca 1080 gcaatgcaac tccgggattc aagcgggaac ggtcagcccc tccaccgcca ccctcacgga 1140 ctttgaaccc atggccaata ggagcgtgac cagcccatgg acgtactcgg ccaatggata 1200 ctatgaacca tccatcgggg aattccaagt gttcagcccg gtggtaacag gtgcctggaa 1260 cccgggaaac atagggatcc gcgtcctccc cgtgccggtt tcggcctccg gagagcgata 1320 cacccttcta tgctatagtc tgcagtgcac gaacgcgagc atttttaatc caaacaacag 1380 cggaaccatg atcgtgggac ccgtgctcta cagctgtcca gcggcctccc tcccgtaagc 1440 gcgccctccc caccgcgtgt caaataaaga gtcatgaacg tt 1482 <210> SEQ ID NO 20 <211> LENGTH: 3510 <212> TYPE: DNA <213> ORGANISM: Infectious bronchitis virus <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(3510) <223> OTHER INFORMATION: Infectious bronchitis virus strain ArkDPI- derived commercial vaccine D spike glycoprotein gene <400> SEQUENCE: 20 atgttggtga agtcactgtt tctagtgacc attttgtttg cactatgtag tgctaattta 60 tatgacaacg aatcttttgt gtattactac cagagtgctt ttaggccagg acatggttgg 120 catttatatg gaggtgctta tgcagtagtt aatgtgtcta gtgaaaataa taatgcaggt 180 actgccccaa gttgcactgc tggtgctatt ggctacagta agaatctcag tgcggcctca 240 gtagccatga ctgcaccact aagtggtatg tcatggtctg ccaactcttt ttgtacagcc 300 cactgtaatt ttacttctta tatagtgttt gttacacatt gttataagag cggatctaat 360 agttgtcctt tgacaggtct tattccaagc ggttatattc gtattgctgc tatgaaacat 420 ggaagtgcta tgcctggtca cttattttat aatttaacag tttctgtgac taaatatcct 480 aagtttagat cgctacaatg tgttaataat tatacttctg tatatttaaa tggtgacctt 540 gttttcacat ctaactatac tgaagatgtt gtagctgcag gtgtccattt taaaagtggt 600 ggacctataa cttataaagt tatgagagag gttaaatcct tggcttattt tgtcaatggt 660 actgcacatg atgtcattct atgtgatgac acacctagag gtttgttagc atgccaatat 720 aatactggca atttttcaga tggcttctat ccttttacta atactagtat tgttaaggat 780 aagtttattg tttatcgtga aagtagtgtc aatactactt taacattaac taatttcacg 840 tttagtaatg aaagtggtgc ccctcctaat acaggtggtg ttgacagttt tattttatac 900 cagacacaaa cagctcagag tggttattat aattttaact tttcatttct gagtagtttt 960 gtttataggg aaagttatta tatgtatgga tcttaccatc cacgttgtag ttttagacct 1020 gaaaccctta ataatggttt gtggtttaat tccctttctg tttcattaac atacggtccc 1080 attcaaggtg gttgtaagca atctgtattt aatggtaaag caacttgttg ttatgcttat 1140 tcatacggag gacctcgtgg ttgtaaaggt gtctatagag gtgagctaac acagcatttt 1200 gaatgtggtt tgttagttta tgttactaag agcgatggct cccgtataca aactgcaaca 1260 caaccacctg tattaaccca aaatttttat aataacatca atttaggtaa gtgtgttgat 1320 tataatatat atggcagaat tggccaaggt cttattacta atgtaaccga cttagctgtt 1380 agttataatt atttatcaga cgcaggtttg gctattttag atacatctgg tgccatagac 1440 atcttcgttg tacaaggtga atatggtcct aactattata aggttaatcc atgtgaagat 1500 gtcaaccaac agtttgtagt ttctggtggt aaattagtag gtattctcac ttcacgtaat 1560 gaaacaggtt ctcagcttct tgagaaccag ttttatatta aaatcactaa tggaactcgt 1620 cgttctagac gttctgttac tgaaaatgtt acaaattgcc cttatgttag ttatggcaag 1680 ttttgtataa aacctgatgg ttcaatttct gtaatagtac caaaagaact ggatcagttt 1740 gtggcacctt tacttaatgt tactgaatat gtgctcatac ctaacagttt taatttaact 1800 gttacagatg agtacataca aacgcgtatg gataagatcc aaattaattg cctgcagtat 1860 gtttgtggca attctttggc ctgtagaaag ctgtttcaac aatatgggcc tgtttgtgac 1920 aacatattgt ctgtagtaaa tagtgttggt caaaaagaag atatggaact tttaaatttc 1980 tattcttcta ctaaaccagc tcgttttaat acaccagttt ttagtaatct tagtactggt 2040 gagtttaata tttctctttt gttaacaccc cctagtagtc ctaggaggcg ttcttttatt 2100 gaagatcttt tatttacaag tgttgaatct gtaggattac caacagatga cgcatacaaa 2160 atgtgcactg caggaccttt aggctttctt aaagaccttg catgtgctcg tgaatataat 2220 ggtttgcttg tgttgcctcc tattataaca gcagaaatgc aaactttgta tactagttct 2280 ttagtagctt ctatggcttt tggtggtatt actgcagctg gtgccatacc ttttgccaca 2340 caactgcagg ctagaattaa tcacttgggt attacccagt cacttttgtt gaagaatcaa 2400 gaaaaaattg ctgcttcctt taataaggcc attggtcata tgcaggaagg ttttaggagt 2460 acatctctag cattacaaca aattcaagat gttgttaata agcagagtgc tattcttact 2520 gagactatgg cagcacttaa taaaaatttt ggtgctattt cttctgtgat tcaagacatt 2580 taccagcaac ttgattccat acaagcagat gctcaagtgg atcggctcat aactggtaga 2640 ttgtcatcac tttctgtctt agcatctgct aagcagtcgg agtacattag agtgtcacaa 2700 cagcgtgagt tagctactca gaaaattaat gagtgtgtta aatcacagtc tattaggtat 2760 tccttttgtg gtaatggacg acatgtttta accataccac aaaatgcccc taatggtata 2820 gtgtttatac actttactta tacaccagag agctttatta atgttactgc aatagtgggt 2880 ttttgtgtaa gtcctgctaa tgctagtcag tatgcaatag tgcccgctaa tggtaggggt 2940 atttttatac aagttaatgg tagttactac atcactgcac gagatatgta tatgccaaga 3000 gatattactg caggagatat agttacgctt acttcttgtc aagcaaatta tgtaagtgta 3060 aataagaccg tcattactac atttgtagac aatgatgatt ttgattttgc tgatgaattg 3120 tcaaaatggt ggaatgatac taagcatgag ctaccagact ttgacaaatt caattacaca 3180 gtacctatac ttgacattga tagtgaaatt gatcgtattc aaggcgttat acagggtctt 3240 aacgactctc taatagacct tgaaacacta tcaatactca aaacttatat taagtggcct 3300 tggtatgtgt ggttagccat agcttttgcc actattatct tcatcttaat actaggatgg 3360 ttgtttttca tgactggttg ttgtggttgt tgttgtggat gctttggcat tattccttta 3420 atgagtaagt gtggtaagaa atcttcttat tacacgactt ttgataatga tgtggtaact 3480 gaacaataca gacctaaaaa gtctgtttaa 3510 <210> SEQ ID NO 21 <211> LENGTH: 1617 <212> TYPE: DNA <213> ORGANISM: Avian metapneumovirus <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1617) <223> OTHER INFORMATION: Gene F of Avian metapneumovirus isolate IT/Ty/A/259-01/03 <400> SEQUENCE: 21 atggatgtaa gaatctgtct cctattgttc cttatatcta atcctagtag ctgcatacaa 60 gaaacataca atgaagaatc ctgcagtact gtaactagag gttataagag tgtgttaagg 120 acagggtggt atacgaatgt atttaacctc gaaataggga atgttgagaa catcacttgc 180 aatgatggac ccagcctaat tggcactgag ttagtactca caaagaatgc tttgagggag 240 ctcaaaacag tgtcagctga tcaagtggct aaggaaagca gactatcctc acccaggaga 300 cgtagatttg tactgggtgc aattgcactt ggtgttgcga cagctgctgc cgtaacagct 360 ggtgtagcac ttgcaaagac aattagatta gagggagagg tgaaggcaat taagaatgcc 420 ctccggaaca caaatgaggc agtatccaca ttagggaatg gtgtgagggt actagcaact 480 gcagtcaatg acctcaaaga atttataagt aaaaaattga ctcctgctat taaccagaac 540 aaatgcaata tagcagatat aaagatggca attagttttg gccaaaataa cagaaggttc 600 ctgaatgtgg tgaggcaatt ctctgatagt gcaggtatca catcagctgt gtctcttgat 660 ttaatgacag atgatgaact tgttagagca attaacagaa tgccaacttc atcaggacag 720 attagtttga tgttgaacaa tcgtgccatg gttagaagga aggggtttgg tatattgatt 780 ggtgtttatg atggaacggt cgtttatatg gtacaactgc ccatattcgg cgtgattgag 840 acaccttgtt ggagggtggt ggcagcacca ctctgtagga aaaagaaagg caattatgct 900 tgtatactga gagaagatca agggtggtac tgtacaaatg ctggctctac agcttattat 960 cctaatgaag atgattgtga ggtaagggat gattatgtat tttgtgacac agcagctggc 1020 attaatgtgg ccctagaagt tgaacagtgc aactataaca tatcgacttc taaataccca 1080 tgcaaagtca gcacaggtag acaccctgtc agtatggtag ccttaacccc cctagggggt 1140 ctagtgtctt gttatgagag tgtaagttgc tccataggta gcaataaagt agggataata 1200 aaacagctag gcaaagggtg cacccacatt cccaacaacg aagctgacac gataaccatt 1260 gataacactg tgtaccaatt gagcaaggtt gtaggcgaac agaggaccat aaaaggagct 1320 ccagttgtga acaattttaa cccaatatta ttccctgagg atcagttcaa tgttgcactt 1380 gaccaagtat ttgagagtat agatagatct caggacttaa tagataagtc taacgacttg 1440 ctaggtgcag atgccaagag caaggctgga attgctatag caatagtagt gctagtcatt 1500 ctaggaatct tctttttact tgcagtgata tattactgtt ccagagtccg gaagaccaaa 1560 ccaaagcatg attacccggc cacgacaggt catagcagca tggcttatgt cagttaa 1617 <210> SEQ ID NO 22 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer pCI-Neo <400> SEQUENCE: 22 ggctcatgtc caatatgacc gccat 25 <210> SEQ ID NO 23 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer EGFP <400> SEQUENCE: 23 aaaaaagaat tcaccatggt gagcaagggc gag 33 <210> SEQ ID NO 24 <211> LENGTH: 38 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer EGFP <400> SEQUENCE: 24 aaaaaagcgg ccgcttactt gtacagctcg tccatgcc 38 <210> SEQ ID NO 25 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer EGFP-I <400> SEQUENCE: 25 gagaagcgcg atcacatggt 20 <210> SEQ ID NO 26 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer beta-actin <400> SEQUENCE: 26 aaaaaaacta gttggtcgag gtgagcccca cgtt 34 <210> SEQ ID NO 27 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer beta actin <400> SEQUENCE: 27 aaaaaagaat tcgcccgccg cgcgcttcgc tt 32 <210> SEQ ID NO 28 <211> LENGTH: 44 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer L2R <400> SEQUENCE: 28 aaaaaaacta gtcatagtaa atatgggtaa cttcttaata gcca 44 <210> SEQ ID NO 29 <211> LENGTH: 45 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer L2R <400> SEQUENCE: 29 aaaaaagaat tcatggtttg ttattggcaa atttaaatta atgtt 45 <210> SEQ ID NO 30 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer Luc <400> SEQUENCE: 30 aaaaaaccta ggttggcaat ccggtactgt tggt 34 <210> SEQ ID NO 31 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer Luc <400> SEQUENCE: 31 aaaaaattcg aagccgcccc gactctag 28 <210> SEQ ID NO 32 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer SV40 <400> SEQUENCE: 32 ggcctctgag ctattccaga 20 <210> SEQ ID NO 33 <211> LENGTH: 42 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer WPRE <400> SEQUENCE: 33 aaaaaagcgg ccgctcaacc tctggattac aaaatttgtg aa 42 <210> SEQ ID NO 34 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer WPRE <400> SEQUENCE: 34 aaaaaagcgg ccgcgcccaa agggagatcc gac 33 <210> SEQ ID NO 35 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer VF1 <400> SEQUENCE: 35 aaaaaaagat cttacatgaa tgacgctgct g 31 <210> SEQ ID NO 36 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer VF1 <400> SEQUENCE: 36 aaaaaaacta gtaaatacac cgagaaatac cacg 34 <210> SEQ ID NO 37 <211> LENGTH: 38 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer VF2 <400> SEQUENCE: 37 aaaaaacaat tgaaataaag actcagaacg cattttcc 38 <210> SEQ ID NO 38 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer VF2 <400> SEQUENCE: 38 aaaaaaggta cccccccgca gagaattaaa a 31 <210> SEQ ID NO 39 <211> LENGTH: 18 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer E1 <400> SEQUENCE: 39 acatgaatga cgctgctg 18 <210> SEQ ID NO 40 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer E1 <400> SEQUENCE: 40 cccccgcaga gaattaaaa 19 <210> SEQ ID NO 41 <211> LENGTH: 496 <212> TYPE: DNA <213> ORGANISM: Avian encephalomyelitis virus <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(496) <223> OTHER INFORMATION: Internal Ribosomal Entry Site (IRES) from a Canadian isolate of Avian encephalomyelitis virus <400> SEQUENCE: 41 tttgaaagag gcctccgggg tgtccggagg ttctctttcg acccaaccca tactgggggg 60 tgtgtggggc cgtatttgga gtgcacggta tatatgcatt cccgcatggc aagggcgtgc 120 caccttgccc cttgacgcat ggtatgtgtc attatttgcc ttggttaaac cccataaaac 180 gaggcgccac gtgccgaaaa tccctttgcg tttcacagaa ccatcctaac catgggtgta 240 gtatgggaat cgtgtatggg ggtgattagg atctctcgta gagggatagg tgtatcattc 300 aaatccaggg agtactctgg ctctgacatt gggacatttg atgtaaccgg acctggttca 360 gtatccgggt tgtcctgtat tgttacggtg tatccgtctt gacacactga aagggtattc 420 ttgggtaatc ccttcctact gcctgatagg gcggcgtgcc cggccacgag agattaaggg 480 tagcaattta aacatg 496 <210> SEQ ID NO 42 <211> LENGTH: 18 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer ORF1-2Ver <400> SEQUENCE: 42 gcacagtccc aatggctt 18 <210> SEQ ID NO 43 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer ORF1-2Ver <400> SEQUENCE: 43 aaattctggt ccgttaccga 20 <210> SEQ ID NO 44 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer TR2-17-11Ver <400> SEQUENCE: 44 gcctactcct cagcctatca c 21 <210> SEQ ID NO 45 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer TR2-17-11Ver <400> SEQUENCE: 45 cagtcctcta acgagcacgc 20 <210> SEQ ID NO 46 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer CAT <400> SEQUENCE: 46 gtgtaggctg gagctgcttc 20 <210> SEQ ID NO 47 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer ORF1CATSwaI <400> SEQUENCE: 47 ggacgtgctt taggtaattt cctccg 26 <210> SEQ ID NO 48 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 48 ttctttttcc cgtttaggtg aggag 25 <210> SEQ ID NO 49 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 49 atttaaatgt gtaggctgga gctgcttc 28 <210> SEQ ID NO 50 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer ORF2CATSwaI <400> SEQUENCE: 50 gcgttctgag tctttattgg taa 23 <210> SEQ ID NO 51 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 51 actcgaaaca cgcgtcacac gcgc 24 <210> SEQ ID NO 52 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 52 atttaaatca tatgaatatc ctccttagtt c 31 <210> SEQ ID NO 53 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer TR2-17-11CAT <400> SEQUENCE: 53 ccccctggcg gccgttggcc gccactg 27 <210> SEQ ID NO 54 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 54 cacccgcgcc ggacttagtc tct 23 <210> SEQ ID NO 55 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 55 atttaaatgt gtaggctgga gctgcttc 28 <210> SEQ ID NO 56 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer TR2-17-11CAT <400> SEQUENCE: 56 agtagagatt ataggaagcg gggatta 27 <210> SEQ ID NO 57 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 57 tagtggtttt tatttaaacg aga 23 <210> SEQ ID NO 58 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 58 atttaaatca tatgaatatc ctccttagtt c 31 <210> SEQ ID NO 59 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer ORF17CATSwaI <400> SEQUENCE: 59 tccccctggc ggccgagggc cgcca 25 <210> SEQ ID NO 60 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 60 ctgcacccgc gccggactta gtctct 26 <210> SEQ ID NO 61 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 61 atttaaatgt gtaggctgga gctgcttc 28 <210> SEQ ID NO 62 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer ORF17CATSwaI <400> SEQUENCE: 62 gatacgaaga aggataggag cagaat 26 <210> SEQ ID NO 63 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 63 cgaggatacg gtagttgact cc 22 <210> SEQ ID NO 64 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 64 atatttaaat catatgaata tcctccttag ttc 33 <210> SEQ ID NO 65 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer EGFPcaSwaI <400> SEQUENCE: 65 agctgcattt aaatgtatta ccgccatgca ttag 34 <210> SEQ ID NO 66 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer EGFPcaSwaI <400> SEQUENCE: 66 agctgcattt aaatccacaa ctagaatgca gtg 33 <210> SEQ ID NO 67 <211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer mCherry <400> SEQUENCE: 67 agctgcattt aaatatggtg agcaagggcg aggagg 36 <210> SEQ ID NO 68 <211> LENGTH: 39 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer mCherry <400> SEQUENCE: 68 agctgcattt aaatctactt gtacagctcg tccatgccg 39

1 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 68 <210> SEQ ID NO 1 <211> LENGTH: 45063 <212> TYPE: DNA <213> ORGANISM: Fowl aviadenovirus D <220> FEATURE: <221> NAME/KEY: whole genome <222> LOCATION: (1)..(45063) <400> SEQUENCE: 1 catcatctat atatacctac atgaatgacg ctgctgtcac ctcgcactac cgctcccgcc 60 cgctagatgg caaaattgct ccaccaaagt atttccgtga tacaaagtcc ggactgggcg 120 cggcgaatac gtgccgggcg aggcacggag ccgactttct ggcaccgata gcaataaatg 180 ggcgttccct tgcgtgcgga attgcgccgg cacagtccca atggcttggg agcggtccag 240 aaagcgatta gtttttgcgt acgcacgcga aagctatttt tgcagcactt acttccttat 300 ttgtttacaa ccgcctccgc ctgtgtgtaa tttaaaattt gtacataccg ttccgatcgc 360 cgtacttggg cttggaacgc ggacttcgag tccctaaatt gatttatggt gtcttaatca 420 gcgctttttg tgccacgttg tgcgttctcc gttgaccttt gaaaaggcgc gtggtatttc 480 tcggtgtgtt tttcgggcgg attacgcgtt acgtgatcgt tttatggcgg gctccgggcg 540 tataacagtg gagcggctga tcgtcagagc acggatgagc gagcagccgg actcaccgcc 600 ggagctctat ccgtctctcc tggtcctgcg ggtgaacccg gttccccgta ctccgttcgt 660 tttggacatc agccgtttgc ccgtggtgcc ggagcgcatt ttgctgcacg actttccgaa 720 cggaccctgg tcggaaacga tttggtacgg gaagatcact cagcgggagc tgaaccaggc 780 gctagagtct atcgtggacg tgctttaggt aatttcctcc gttctttttc ccgtttaggt 840 gaggagatgt ctttcgattc cggttgcccc ccgactccac cggtaaagct gctgttcaag 900 aagcattccc cgtttgccgt gacgccccag cgtgctacct cgggggctgc cggttacgac 960 ttgtgcagct cggcggacgt ggtggtgccg ccgaagagcc ggtctctcat tccgaccgac 1020 ctttcttttc aattcccccg aggcgtgtac gggcgcatag cccctcgttc gggcttggcc 1080 gtaaagtttt ttatcgatgt gggagccggt gttatcgata gcgattaccg gggcatcgtt 1140 agcgtcctcc ttttcaattt ttcggaccac aatttcaacg tccgtcgggg cgatcgcata 1200 gctcagttaa ttttggagcg tcatttgacg cccgacttgg aagagcgatc gggcttagac 1260 gagacggcca ggggtgcagc cggtttcggc tccaccggcg gttttgatac aggggtctgt 1320 ccctcctcct tttcttagga attgaccgag agcgaggcgg cggacgaagt ggatttcgcc 1380 gttcaactgg cggccgctct cgaggcagat aacgaggcta ctgcgggttg cttctgcgaa 1440 gccgactccg ggaaaaacac ctgttctctg tgttcttgga actgtatcat tctttaggta 1500 tgattgtttc tgtttggacc ctcgttgtag agctcgttaa cgcgtgggat cgtttgtgtg 1560 gtcgtcgggc ttaccgctat tacgcggatc gcgagctgtt gtggcaagtg cacgagagtc 1620 tgcggtccgc cttggagaag acggtcggtg tcggaagagg cttggcccgc atcggcgacg 1680 gtgaaaacgg gtatttcgga ggaggcggcg aagctgttcg cgacaattga cgacatgtcg 1740 atcgaagatt ggttcctcgt tttcgtagtg cttttcgcta ttctgtcttt tatgtggggg 1800 atgttcttgg ggatgctttg cgggccaggc gatttggatt tcgcgggtta ctgggactgg 1860 tatttccgtg tttgtaacgc gagggttcct taataaatgt cttttcgtaa cgttaattcc 1920 gcgtttgggt tttgcttttt gcagacggga tgtcgggctc gagggatcgc tattggtcct 1980 tggtcgaatc tctgataaat agagggatcg tgacccgtca gcagtggtat ttcaacgacc 2040 gctccgagta cgggtattac acgaagcgct acagaggcgc gatgaaattg tccaacttgt 2100 tccgcgacgt gcgtcgtcac atgttctgga caaaatgttt ggccgactac atgttgaccc 2160 ccgaaaccga ctttcttcaa atcgaggaga atccgtttta ttcggtgttt tcgaaaaacg 2220 gctacgatcc cgctctcgtg ggctcgatgt tgctggcttg ggccgcgcgg cagtcgaagc 2280 gcaataccgt ttggattagg ggagcgcccg tgacgggagc tccttacttg gcggaagcga 2340 tcgcttactg ctgcccgttg atgacgtgcg tcgattggcg caacaagcag aatccttttg 2400 agaaggggtt cgagtctttg ctgttttggt gggacggcgg gcacgtgccg caccgctgcg 2460 tggccttgtg tcgccaggtg tttaggggcg aaaacgtcat gtttccccgg gaagacggtc 2520 aaatgagcga attggtgcgt acgcccgtgt tgttgtactc tgaacacgac gcttgtcgca 2580 cgttagtcaa gtggggcgtg ttcagtgatg agttttctga cggtttacgt aattccatgt 2640 attgtttaga cttgtccgcg cccatggaag gcgctcagtg cgtcacttgt gccgacgctc 2700 gcgcgttttt tacgtgggct tacccgcgac gcatggattt tgactgtttt tgagcgcgtg 2760 tgacgcgtgt ttcgagttta ccaataaaga ctcagaacgc attttccgtg tgaccgatgc 2820 tttttattgt taaatttcaa cgggaaagct tgcaaaacgg atagcatatg acgactagcc 2880 taaaggggaa ggtttcgagg ttgtcctgga cgcgacgaca acagcgcatt aagctgttgt 2940 taccgtagga gggagtgatg aatatctcgt aaccgcctcg tatgcgggat aaggcgtaat 3000 tattgcggac ggcgcggtgg gtcagcaagt cgtgcagagg ggccagaggg ttcaggatgt 3060 agtcgagtcg tcccgtatcg tcgtcggggg cggtggagat gagtaagagg ttgaaattgg 3120 tgatgttaac gagaggatag gggttgttcg cgcgctcttc ttcaaagtac tcgtacactt 3180 tatccaacca gtatgtgcac acgactttgc tcgctgcttg acacatgagg ctgtagggat 3240 tggtgcaggt gcaatcgtat cggatttgag gcaaggggga ttgctgctcg aggtcttcgg 3300 taacggacca gaatttattg tacagacgac agcgcgccct gcatttgggc atgctgtatt 3360 cgggcagtat ttcttcgaag gtgacggtgt ccttcctggg gaacggaaac atgcgttctc 3420 cgtgaagaaa ctctaagaaa aagatgtcaa gagcgaggtg agcgggtgct tccagctggg 3480 cttctttgca ttctagcgtg agagagtgca cgctgtcctg cggaaagaga gaaaattaca 3540 ggaacacggt gatgacgggg tacgggaact tttgcatgag tctcaggaag cacggtatga 3600 gtgtgtaatc cctgtgcgac cgcggtaacg cgatgcgaat gtgtccgtcg accacttcta 3660 tcttggcgaa gggtttggag gaatacacga ataggttgat gacgccgtcg atgatttcga 3720 tgggaggtct gccgtgacat cggaagcagt gcgtgcaacg ttttcggacg tattggtcga 3780 aatggatggg aaacgggttt tccgcgatgg ggtacgcggt gagtttggcg tgccactcca 3840 tggtgagtgt ggcggctagg gactgacaaa agagcgagag cggctcctcg cactcgcata 3900 cgtagctcgc gacccattct cccatccatt tccgacctcc gtcctctacg tcggctaata 3960 ggttgctaat tttaaaggag gactggaacg gcctcagcct gaacggcagg gattcggggg 4020 taaagtaggg ctctgaccga tagggataca tggatcttcc gcataggaaa cgcagtagat 4080 ccggatcgaa aagggccaag gccgacacga cccgcagggg cgaggggaat tttacggagt 4140 agacgtgttc gcaggtgtca ccgaggttgt cgctgtccca tcttatgtga aaagtggacg 4200 aataacgaga gggagacgag atctgcaaag aaaagaaagt tttattgggg gataactttt 4260 tcagtattcg ggagagtagc ccgcgcaagt ctgacaaagc ctgttttcgc gcttgcagtg 4320 catgaccgct gcgcaaggta cggttctgga ggtcgacgcg acaaagtctt ttaattctct 4380 gcgggggttg tggcagacgg gggtgagctc ttggggagcc gtgaggaaca ggatggtcga 4440 gtttctgcac aggaacgaac aggcgataga gacgtcggtg caatggatga gcatttgggg 4500 cgctctgacg aagacggggc gaccggcgat gtagcaggtg cccgaattgc cctccataaa 4560 gtgtagcgcg agcgtggacg gggctctgtc gacgaagggc acgcagtaga gcgacgtgtg 4620 ttgggcgacg ttggctatcc gctctatgtc gttgagatcg ctatctgaga tggccattgg 4680 gaagcaggag gcgaggacgt tgtacagcag cttggcgttg gcgaggcgtc ctccgcacag 4740 gatgagcgta ttgatctctc cgttgagcca ccgtttgatg gtgagggcgc acagagccgc 4800 gtcgtacccc tctttttcga gcagctgatg gaaccagttc tcttcgggat tgtagaaagc 4860 gagtctgagg gactcgggtt cctcgtaggg cagttgggcg tagagagatc tggagtttcc 4920 gaatctttct cggagggccc ctaggacgta ttcccgctcg tccggtctga gggtgtactg 4980 gtcgtaggcg ggcatctggt cccactgaga tcggtgagcg atgccgtctc ggaaaagccg 5040 ttcgatgagc gcgtccatct gcgacgccga cctgttggtc tcctgagagt agcgggaagg 5100 agtctttttg ttcggatcgg gtgtgggcgg agtttctggg acgtaaggaa agccggagaa 5160 gtcgtcttcg tcgtcctccg tgcgcctctt ctgcgacggg ggcggcgagg agggagaggg 5220 actacgatgt tccgctttcc gcttactctg gaagagagag gatatgatca aacttctgct 5280 tgcaggagta acacggggga tcgatccagg cggtggagca gagcactccg aaatcgtttt 5340 tgcagtcaaa atcgcgttcc ccttcgtgct cgcgtatctg cctcaggtat tgcacgagct 5400 cgctggtgag gaaggagatt ccgtcgtcgt tggcggtttc gaactggatg atgaggtggt 5460 actttgggtt ggtgggtagt tggtctgggc gcggcagctt gttgagtcgc acgacgcact 5520 tcacttcttc gatttcgacg agttcgccct gatgcgagac gttgaaagcc ctgcccctga 5580 gcatttcgtt gacgagtggg tttttaaagg ggtactcgtg atgcaggggc gggaagtaga 5640 tgagtttagt gacgtctcgc acgggaacgt actggtcgaa atcgaagttg ttgacgtgcg 5700 cggtcaggac gcagtggaac atgcgcaaga tagagttggt aaaggtgtct gcgcaggtgg 5760 aggagtcgcc cacaatgtag agaacgttga gagtgtcgtg caagcgcgaa agcgtgtagt 5820 cgctgagcca gcgaccgagc atgtagctta gggtgacgag attgaaccct tccctttccg 5880 cgaatcttgc gagatgcctg ctggtttcta tggtgaaggg gccgctaggg ctattgagta 5940 gggtttggta gaggctaaag ttgcagtaga ttttcctggc ttcgtggaga agcagttttc 6000 cctgcagacc tactccgtat cgagcgaatt gctccggatc cctgagacgc cattctcctt 6060 cggtgaaaat gagcctcttt tggagctggt aggcgagttc gcccggattt tcgggggcgt 6120 tcgaccgctt attggaaccc gtttgcctcg acctgcgact ctggagggga agacaggggt 6180 tcgaagacgg ttagtttagg gcgcttggcg agcatgtttt tgcgagcctc tgcgaatcgt 6240 atgatgagct tggaggctct gaagaatttt tcctgcacgt tgatgttaag gggcaggatg 6300 gcgtaccgct gatcgagcac gcaccagcgg aacgagtcgg agacggggtc gggggagtaa 6360 gtgagccagc tgaacttttg gttctgctgg agaaaagcga agtaggcttt gagtaacacg 6420 accagttctt tgctgatgtt gtgggcgtaa gagtagagaa agcgcgagaa ctgaaattgg 6480 gggatgtggc aagagaggat gtgtattttg gcgttgacct taagggtcgg cacgtttcct 6540 atggcggtgc gcggcgcgag attgtggaga accacgaaaa cgtagaaagc ggtacagtta 6600 gcggaacgag cgaagagctt agaggggaga gcgtgaaaga gcacggagac gctagacccg 6660 gagcagagtt tatccatgca ctcgtccatg acgatggcca gcggtccctg ccgggacgcg 6720 ttgacgtata cgttgtccgg gtgttcgatg ttgaggtttt cgggcgaggt tgcttcctcg 6780 tacgtcattt ctacgaactc cggtcggaag gtgcaggttc tgggcgcgaa ggttccgtcc 6840 tccttgcagt cgtagttgga ttccagaagt tgtaggttcc aagccgtttg ttccaccggc 6900 gggatcatat tcttctccgg cgtcacaaag ataaccgttt ccgggatcgg ttgcagcata 6960 tcgcaggaga tcaacgctct gagcagatgg ctcttgcccg atcccgtcgg gccgtagatg 7020

acgccgatga cgggctgtcg gcccatgttg agcgacggta gttgtccgtc gacgaggtac 7080 cgactgtctt cctgttcttg ccgttccacg gctagctgta tttctaggaa cttgtcgtcc 7140 actcccccga gactgtaaaa gtcgtcgaag ctgggaaatt tctgttcgcg aaagagatgt 7200 ggggctaggt ccgccgctcc ttcgtaccag cccgttaccc gtttgtagaa ctcgcctgcc 7260 cgtagatact cttcctctcg gtagcgccat gttttcctct ttttattagc gacggccccg 7320 gcctccgctc cgctcatggg tctccgagga gatccaaaat ttcgtcacag tcctcgatag 7380 agagcgaccg gtctgggacg tcgaactcgt ccgagtcctc tccgtcgtcc tcgggagcga 7440 tccgtgcggc cgccgccgcc gcgagcagcc tgaggggggc tagaggttct tcgtggccca 7500 ccgggggcac tctccggact tcgtcggcgg ttctcgggtt gggatgttcg gtgtcgtagg 7560 ggtaaagaaa gtccccgtgg cgatactgcg tgaggtcctt ccagggtctt aggactctgg 7620 tgagctgttc gttgtggatg gtgaacggat cgtatcggct caccttgttg aggagcgtgg 7680 ttttgaagat ggtcctgcgc gtgtgaagct cgggaatggc gctgaccgcg ccgtacgact 7740 ggtcttcgtg cctctgccag cagcgttcca gcgtgtcgta tataagctcc gcctgtctgt 7800 gtcccttgga gcgtattttt ccgcttccca cgtgaccgca ctcggggtcg agacagacgg 7860 cgtctttgag tccgtagagc ttgggggcga ggaaaatggc ttcggagctg tacgtgtcgc 7920 cgccgcaccg cttgcacttg atgtcgcagt cgcaggccca gtagagcgcc gggttctccg 7980 ggtcgaacgt gagccgtgtg tgcgggccct tgatgcggtg cgcgcccctc tccttcatgc 8040 gccgataccc cgactcggtg acgaagaggc tgtcggtgtc cccgtagagg gtctgggggt 8100 ccctttggag taggtgtgtc cccctatcgg gtccgtgcag gatttcgcac cactcgctga 8160 agaaggcgcg cgaccatccc aacacgaagc aagcgatttg cgtggcgtat ctcttatttt 8220 cgaccatttt gtctaaattt tctagatgta gaacggtcaa cgcttcgggg ggagcgtcga 8280 gcaaccgcat gggtttgaaa acggtctcgt tggcggtggc gtaatgggcg tggtcatcct 8340 cggctatata agggggcgcg tcgagcgctt cctgcagttg gtgatcgact tccgccaggg 8400 acgcttcgcg agcggtaagt gcgttttcga gcttactggt actctcgcct tcgctctctt 8460 ccgtttcctc ctcctcctgc tcttcgtttt cttcttctgc tgtccgcgtc gggggtttcc 8520 ggtcaaagta ctgacgcagt tgatgatgtc tgaaggcgtc gggggtgctt tcggggcaga 8580 cgaccgtgcc cgctagcgag tcgtggttga gcaatgtcac gtgacgcacg atttgagtgc 8640 cttcgtagat gccccgcctg tcgtcctcgg tgaggtcctg ttcgaagagg atgcgcgtgg 8700 tgtccatgtt ggtggcgaag gcgccgtaga gggcgttgct tagcattttc gagatggacc 8760 gcataacttc gtttttgtct cgatccgctt tctctttggc cgcgatgttt ttgctgacgt 8820 attcagcgca gacggttttc caagcgggga acacgatgtt catgtcgtcg tggagggccg 8880 tgaccgccca tcctcggttg tgcaacgtga ggatgtctat gacggtgatg acctcgtcgt 8940 acagcgcttc gttggtccac acgaggcgcc cccctcgcct ggagcaaatg ggaggcagcg 9000 tgtctaagtg ttccgcgggc ggaggataag cctcgacttt taggatgcac ggtttgattc 9060 ggtggtcgaa gtagctgatg ggtcgttcgg aactcatgag ctcgttgagc tcgtcgacgt 9120 gtttggcagt gaatttgggg tccaagggca tgccgtgggg catggggtgg gtgagggcgg 9180 aggcgtacat accgcatatg tcgaaaacgt atacgggatg tctgtacggg cctagcacgc 9240 tgggataaca ccgcccccct cgcaacgctt gccgtatgta tttgaacatg ggacggtgcg 9300 gggcgtatat ttcggcgacg tactcggggg agggaagcct cttttttctt tttttggtgt 9360 ttttgtttgc gcttgcgcgc tcgacgtcgt gtacgtcggc gtcgtgaggg ttccccttga 9420 tgtcgttttt cttgagcgcg acttttcttt tgtacttgga aggggggcgc tcgaccgtgc 9480 tcgcgcgttc ctgctcttcg acgtacgcgg tgaaggtaag ttgcttccaa aaggcgtggg 9540 tgttgctggg aatggtgggt cgcaggaaaa tgttaaagtc gccgtgcatg ccgagttcgt 9600 ctttgaagta cttgtcgtaa ctgtcgtaca aggtgagagc taatttctgc gtgaccctta 9660 cgtcttgcat gcagtactcg aggcacgcct gcacgatgtc gtacggacga cccgggtgct 9720 cctgtcgcca tagcgccttc tgttcggcga tgacagaagg atcctcccag tatctcgcga 9780 cggggaaccc gtcggagtct tgttcgtaac cgccgatgga gacgtgttcg ttgacggcct 9840 cgtaggggca ctggcctttg gagagctcta gggcatacgc ggcggcggct ttggagagtt 9900 tggctccgct ggtgaactgc aacgtgtcgc gtaccatgaa tcgcacgaag acggtgcggg 9960 agtcgctggg cgcgacgact cctcgcgccc acctgtgaag ccgcgtgggg tcctttttat 10020 cgtagttggg gttgggcagg tggaagagga cgtcgttaaa gaggaggcgg cctacgcgcg 10080 gcatgaagaa gcggtcgcac cgacacgcct gcggaaagac gtcgcgccgt tccacgagtt 10140 cggtggcgag caagagctcg tcgaacttgc agatgttgtg tccgagcacg acgatgtgaa 10200 cgttgttaaa gtcgtcgggc agtttgagag gttcggtagg tttgtcgaac agttcgtagg 10260 ggatgtcgta gcgggaaagg tatttgcctt cgctcatgag cttgctgacg tagtctccgt 10320 tggctcgaca gtacctgtcc acgagatccc tggcgaagtg ttgttgcaaa cgggaccggt 10380 aggcgcggaa ccgccgggca acgaagccgg gccgcgtgtc tagccagtag aagccctctt 10440 gtaaagtttt tacttccggg tcaacgagcg ccaggtccgt ggccgcgcgc acgacgtcgg 10500 gatcgcccgt gagcatgaaa cacaacataa aggggtgcat acgttttcct ttattttcga 10560 agacggtata cgtctcgatg tcgtaggtga gaaagagttg cttacagtgg ggttgctgag 10620 cggggcatgc gaaacgcacg tgttgccaca ggtcgcagcc gaatttttgg accgtgtggt 10680 agaaaaaggc ggaccgcctt tcgttgcacg tgtgattgcg cacccagtgg cgtccgcagg 10740 cgggacaggt ttgcacggcc gtgcggctga gtatccaaac ccagtcgccg ttctggtctt 10800 gggtgaccag catgggggga agaaccggat ctccgcgttc cgcgacgcgt ttgacgtgcg 10860 cgagacgccc tcggaagcgg acgacgccga cgtaagcggg tttgaagacg ggcagtcttt 10920 ccggggggtt ttcttggtag tacccgtagt ctattctatc gtatccgcgg cacctgcgcg 10980 tgcctgccgt aggaacttgt gtagtcgcaa gaattttttc atgcctttgt ggaacacggc 11040 cggttgcaga cggaacggta cgccgtcgat gtagtagacg ttggcgcgga tgggatctcc 11100 cttggccgcc ttgaagtggg tcttggcatc gttttggctg agcgtcttaa ccaacggcgt 11160 gtcttccggt ccgagcagag tgttttctag gggaggcggg gagtacgcgg cacgggaaga 11220 ggaggaggca gacgacgaga ggctcatcgc atgcgaggag gcgcgtgccg agcgacgagg 11280 agttcctgtt ctcggtctct caggacccga cgcaggttgt ccgtgatggc ctcgttggtg 11340 ctgatggcca cgatgccccg gaatttgatg cgaaaactga tgtcgatgga gtcgatgaac 11400 tcttcgctga gattgagttg tttgaggacc tcttcgatgt caccggaacg gtctctgtat 11460 tgaatatcgg agagaaataa gtgttggtct gcttcgtcca tgccttcgaa ctgccccgtg 11520 cgctctatca tcatgagaaa atcgcgcagg atgcggttcc acagggtttc gaaaatggtc 11580 gcggggttag attgttcgct ccagatgcgt ttgaaaatag gctctgcgtt gacgtcccac 11640 cccacgatca gtacctgcag ggtggtgacg tctacgtacc gccggaactc gcggttcgcg 11700 acgaagtgcg agtaaagata gtagagcgtg gaggcgacgt gttccgcgag gaaaaagtag 11760 agcacccact ttctaagaaa cgattcggtg acgagcgcca cgtcggcgtc tcgcgtttgt 11820 aggagcagtc ggtaaaagct atccgtgaat cggaacagct cgtgtctgcg ggcggcgccg 11880 gagagttcgt tttgcagggc gtcaatggct tccagcacgg tgcggatgac ttcgtcgagc 11940 agcgtttctt cttcctcttc ttcggggggc atttcttcct cctcttcttc tgggaaggct 12000 tcttcttcgg gaggggccgg gggcggaacg acgcgtctcc gtctgatcgt tcgcggaaga 12060 cgatcgacga attcgcgtac ggctctccgc ctgatgcgcc tgacttgcga ggcggtgatg 12120 gctcgccctt gtctgtccct gggtcgtagc ccgcgtctgt cggggacgcg tctgttgcgg 12180 aggacgatgg cgccgcccga gaagggatcg cctccgtccc cgccaccgcc ggcgcgcccc 12240 gcgtaccctc tgcccgaggc cagagtgcta acgaaacatc tcgccatggt ctccgcaggt 12300 tcctcggcgt tcactctcgg atcgagtgcc gcttgtcgga tcaaactctg cgtgtctaga 12360 tcggcgaacg ccgcttcgaa gagcgtgagc cagttttctt cggcgaacac atcggatcca 12420 ggtaagaacc tatacgtgtt ttccgtatta aaatgttcgc gcgcgtaaca gaataaatag 12480 tggcacagag cgactcgcaa acgccgtatc gcttccaata aggtcgcgtc gtctctcgac 12540 gccgcacgca tgataatgtc gttttgcaaa cctcccgcgc cgctacccga gaggtggctt 12600 cgcgcagcag cgagcgcggc gtccccgtcc ttcccgacgc ccacgacctc cccggagcgc 12660 gcgacgtcga accctctgcc tcgcagatga acgtgcgcca tgtcggaggc gattctgtcc 12720 ataagcacgg cgttgtgcat ctgctgaaag ctgtcgtgaa agttttcgag gtccaggaac 12780 ctcatgtacg cgcctacgtt gacggagtac gagcagtcgg tgaggcacgt ccagaaaagc 12840 ctcctgggtc gctgcgcggg cggactttcg tagccgagtt gcatgaagac gcgattttcg 12900 aaaaagtaat cgttgagcgc gcggtgcatg tactggtagc cgagcaggag gtgcggcgga 12960 ggcagtcggt gatacgggta tcccgcgacg ttgggtccgt tcgggctgag gtccctgagc 13020 tgcatcaatc tgtagtcgta gagtcggctg acgagaaaga cgctgttggg gtgtaccagc 13080 gccggctgat tcctgaccat gggaaagtcg gacgcgataa cgggttgaca aaatcgaacg 13140 gtgtttaagc tttgaccggt gagctccgcg aagactctgt aagcctgaaa ttgagccctg 13200 acgtttttag aggagggcct gttcccaacc gagagacgcc atgcacccgg tactccaaaa 13260 cgttcggaac gcgagcgcgg gcgccggcgg ggaaggacca caccgtcacc agcatcagca 13320 gcaacacgtg caacgccacc accagcaaca gcaacggcac cgacagcagc ttccgcaagc 13380 ggcgccgacg cgaaaccgcg tccgccctcc cacccctccg cagtacccgg cgcagcacgc 13440 gctgccctcg tccgcttccg cggacggctt cgcgtccgac ggcggcgagg acctaggtcc 13500 tcccgtttgc gggctggcgg cgggagcgga cgtagacgag attcgaatgc gaaagcggga 13560 aagcggtcgc cgcggagcga ttcccgagac gaatttgttt aaggcgagca gagaggcggt 13620 cccgcaaaac gattacgata gggaggcgat gtaccggtcg gggcaggcga tcagcgttaa 13680 ccggggtcgc gtgctgaccg ccgacgactt cgcgtacgat gagggacagg acccgtcgtt 13740 cacgccgggt gcgaatcact tgcgcgcggc cgagcttgaa acgcccggcc gagcagaccg 13800 cgttcgggaa gagctgcgca acagctgcca ccaaacgcgt ttgcgtacgg cgttgacccg 13860 tcccgagctg cccgcgggga tctactacct gtacgacttc gtgcagacgt acgtcgatca 13920 tcccgacggg cgagttaagt taaacccgca gctggtttta gtggcacagc acgcgggcaa 13980 taccatgctc gcccaacggt tgtggtcgat agcggaggat aagaacgcgt ggttgcgtga 14040 cctgatcgag atggcgtaca tgatcgtgaa cgatccttac ttgagcaccg agcagcaatt 14100 gtcggcgata tgcacgaccg tggtggagtt gagcatgaaa tacgctaaga tcgcggcgac 14160 gaacggttat ccgtcgatgg cgcagatggc taaggcgcaa gaattctttt accgcgtgat 14220 ggaagccgtg ttagacttgg gagttcaggt tggggtgtac aacaaccgtc cggtgcggtt 14280 ccgtcagaag cgcatcggcg agatccctca gatgtctgac gcggactaca tgttcggttt 14340 gacccaggcg ctcgagaaca ggcctcctca gggcgaactt ttcccgtcgg acgagggaga 14400 atccagcggc ggcgaggcgg aggaggacgg ctacgatgac gactactgag tcgtttatgc 14460 ttttggcccc tctgggtcgc gaggaagtgg cggacgcctt aagctcgtgc cggacgggcc 14520 gacggcgcgt tcgctttcgc catgctccgt actcgaaccg tttgcttaag ttgcaaacgg 14580

cgatggttcc gccgaagata gacggtactt cggaacgcgt ggccgaagtg gttaggggtt 14640 tggccgagca gggcgccatt tatccggacc agatgggggc catacattcc gatctgctaa 14700 accgcgcgta cacgtggaac tcgatgggag tgcaagagag catccaggcc ttggtgaacg 14760 acgtgattca cgggcagaac aaggcgctgc aagacgagct ttcccgtacc cgcgagatcg 14820 cgaacgcgtc gctcttgacc caatttttcg atagcattta caaaacggtg gacaggggcc 14880 agcgtaactt tgagggattt aaaaagctgc tgcggctttt cgttaacaac gtccctaacg 14940 cggaggtgta cggctcgggg ggttcgttca gcatgcagat caacctcgga ggcacgagcc 15000 agaacatcaa tttgactaac gcgttcgaaa acttaaagga catatgggga gcgcgttggg 15060 acgcggtaaa caatccccgg ataggagcgc tgctcacgcc caacacccgg gcgttactgt 15120 ttttcgtgag cgccttctac gattacgggg ctatggagcc cggcagttac ctggataaca 15180 ttctgagatt gtacaaagag gccattaggg cggacataga cgcggaggga gacgccgtga 15240 tggagttggg cgaagcggga gccaacctaa atctgcagtt caacgagtac aaggataccc 15300 tgaactacct gcttcagaac agacagtcgg tgccggatac ggcgccgctc gaattgagcg 15360 aagagcagga aatgcttttc cgctatctga tgcgtcaact aaggcaagcg ctgaaggaca 15420 ggattccttc ggacgtttcc gtgagcacga tggcccaata cgtggacccg cgcctttatc 15480 agacgaataa ggtgtttatc gaaaagcttc aaaactacct gttggccgcg cacgcccgca 15540 atccccatta ctttagatcg atcgtgatgg atcgagactg gcgtcctccc gccgggctct 15600 acacgggtaa ctacgtcata ccggaacgcc tgccgttcga ttattacgac ggcgacagcg 15660 tagtcggagg cccgtcgcga gacgagtact tcgaggagta tccctacgcg agcaaaccgt 15720 acaaaaagat gagcggtcgg gagagggccg actaccagga gcagctggat aacttaacca 15780 cgcagataga tcaggctttg ggcgtgcaat cggaggccgg atggttagcc gatcaccgct 15840 tgccccaagc gttcgatggt gctttgagtc tagcggatct ggcggcgatc ccgttacccg 15900 ccagtccctc cgatcaatcg atggcttcga gccggcgtag ctccctgtcg agcgtaggcg 15960 acatgatgtc gcgcctgaac ttatcggggc gagggggagc gggatttttt gcgagcctca 16020 gaccgacctt aggaagccct agaccttcta gcagcgccag acctagcgga ttagcgttgg 16080 gcctgagggg cacgggtcct gcgcgcgctt acagaacgcc tcgcgtcggc ggcatgtcag 16140 gctccggatt ggctagaagg gctttggcga cgcggtctat gcgcgacggg aaaaggctgc 16200 ggttttacta gcgtagatgc gtagaaacgg aagacgaaga aggaagaatg cgcgaggtgt 16260 aacgagggaa cttaccgcca tttctagagc gatgcttggt caccacgact ttggtttccg 16320 cgaagcggcg ttgaaccggg gttcctcttc ctccttcgcg acgtcgatgc cggccgctcc 16380 tcctccttcg cctgtgtcgg gcgtgccccc tgcgatggct cctcaacgct acccggcgac 16440 ggtgcacgga taccctccgg tgattccgcc actagactcg gccgcggacg acacgtccga 16500 actgttcgtt ccggtgcaga gggtgatggc gcctacgggt ggaaggaaca gcataaagta 16560 ccgcgattac gcgccctgtc aaaacactac caaattgttc tacgtggaca ataaactgag 16620 cgacatcgac acgttcaacc ccgaggcgaa tcacagcaat ttccgaacca ccgtgatcca 16680 taatcaggat cttgatccgg ctacggcggc gaccgagacc atccagctgg acaatcgatc 16740 ctgctggggc ggggaattga aaaccgccgt gaaaaccaat tgcccgaacg tcagttcgtt 16800 tttccaaagc aatacggtgc gagtaaagtt gatgagttac cgcgatccgg tgcctccagg 16860 cacggcagct cccacgagcc ccgagcccta cgctccagcc ggtgctcagt acaagtggta 16920 cgacctaacg attcccgaag gcaattacgc cttacacgag atcatcgacc tgctgaacga 16980 gggagtcgtg caaatttatt taaaagaggg acggcaaaac aacgtgctgc gctccgacat 17040 cggcgtaaaa ttcgatacgc gttactttaa cctgcttcag gacccggtga cgggtctggt 17100 cacgccgggc acgtacgtgt ataagggcta ccatccggac gtggtactgc tgcccggatg 17160 cgcggtagac tttacttaca gtcgactaag tttaatgtta ggcatagcga aacgcgaacc 17220 gtattcgaaa gggtttctca ttacttacga agatctcgag gggggtaacg tgcccgcact 17280 gctagacgtg gaatccgcgc agatgaccgg cgtcgatcaa gacgtgatcg agctggcgga 17340 cgcgaagccc ttgttaaagg actcgaaggg cgtgtcctat aacgtcattt acgactctaa 17400 caaccgtccc gtgaccgctt atcggtcttg gcttatcgcg tacaaccaat cgggttctac 17460 ggcaaacgaa acgaccttat tgaccgttcc cgacgtgggc ggaggcatag gagctatgta 17520 cacatctatg ccggatacgt ttgtggcgcc taccgggttc aaggaagata acacgacgaa 17580 ccttgccccg gtggtgggca tgaacctgtt ccccgccctc aataaagttt attaccaggg 17640 cgcgtccact tacgtgcaac aactggaaaa ttcctgtcag tcggccacgg ccgcctttaa 17700 ccggtttcca gagaacgaaa ttcttaagca agctccccct attaacgtgt cagccgtgtg 17760 tgataaccaa cccgccgtcg ttcagcaggg ggtgctcccg ttgaagaatt ctttggccgg 17820 cttgcaacgt gtgctcatta cggacgacca acgtcgacct ataccctacg tatataagtc 17880 cctggctacg gttcaacctc gcgtgttgag cagctccact ttacaataaa acactcaaaa 17940 atccaaaatc gtcgccgatc gctgaaaacc atgtccattc tgatctctcc caacgacaac 18000 agaggttggg gcatgcgtcg ccgctcgtct atgcgtgggg tcgggattcg tcgtcgacgg 18060 ttgacgctgc gtaccctgct cgggctgggt accagcagcc gacggagaag cggcggtcgg 18120 tccagcagac gccgttcccg ccccgcttct accactactc gcgtaatgct ggtacgtacc 18180 agcccgaaga aggagacgtt aacgcgcgag gtcgactgga aaacgcagat ggaaggagga 18240 ggaatgaaaa attgaaggag gaggaggacg aggacccgcc gttcattttt cttgactgtt 18300 gttttaacac tcgtaatcgg ctgttacgcg agaggcttgt aaataaatgg agagaaaatt 18360 tgcgcaacgg aatgagaaaa gactactgat ttgcctgtac cgtcgctgta ctacgcagga 18420 agcgtcgcca acatcatgcc ggcagtagtg cttacgggcg ggcgaacagc tagagctaat 18480 cggagggcgt caaccgcttc ccgccgaaga ctccctgctt ccaagctacg ggccacccgt 18540 cgaacgcgcc gctcagtcaa cggtagtaag aaaaagcgtt cgccgaacgt ggctgtggcg 18600 gtaccgtcgc ccactacggc ctccgctgcc gagcgagccg cgctgcagaa cttagctacg 18660 cggttacagc gagggcagtt tacggcttgg cgttcggcga actaccccgc ccctgcggcc 18720 tccgaggccg ccctggccgc tgctcaaagc ggcgcacccg ctacggccag ggacatgacg 18780 accggaacga ccgctacggc ggttcccgtt tcaggtgccg ggatcgcgtg ttcaacgagg 18840 agaaggagaa acggtgttag cggcggtaga acgcgtcgcc ggggaaaaca gctgaaaggg 18900 ggaattttac ccgctctcat acccatcatc gcggccgcta tcggggcgat tcccggaatc 18960 gccggcacgg ccgtcggcat tgccagtttg aaagaacaac agagacagtt caataagatg 19020 tacaatcaga agtaagagag agaaaagaaa aaaaccgcgg gtgccggtgc tgtctaaaag 19080 gccgttttca ataaaaattt ttatagaacc gctgtcgccg ccgtcgtttg ccgtttgtcg 19140 tcgtcgtcgt cgtgagcagt tgtcatggat tacgccgcgc tttctccgca cgtcggtggt 19200 tgggccctgc gagacggata cctcggagac tctagcctgc gagggggtgc tatcaattgg 19260 tcgaacgtgg gctctcgtct ctcgagcgct ctcggcacgg ccgggcgatg gatgtttaac 19320 cagggaaatc gctttctcaa ttccaacacg ttcggccaga tcaagcaggg ctttaaggac 19380 agcggagtga tccgtaacgt cgcaaacctg gccggggaaa ccctaggcgc tcttaccgac 19440 ataggtcggt tgaaaataca acaagatcta gagaagcttc gccgtaaagc cctgggagag 19500 gacggacccg ccactcaagc cgagttacag gcattgatcc aggcgttgca agcccaagtc 19560 gccgcggggg aatccgttcc cactaccgct cctacggtag ggccttcggc tcccgccgaa 19620 gtgcctcgac cgacaacacg ccccatcccc gagatggtca cggaggtgag gcctcccgtg 19680 acgtcctcgg ctcccgcggt gcccgaaacg gaccggccga cgaccctcga aatgcgtcct 19740 ccgcccgcga agaggaggag aaagaggcca gcccccggat cgtggcgtac tcgcctaaac 19800 agcctttcgg gaaacggggt cgcagttagc cgccgtcgca tgtgttattg acctctcgat 19860 aaaaaagcgt ggagatgaaa aaggcattaa atcgtatcta tcggattacc aaaaatttag 19920 catgtcgccg tttttttagg tcgatccgcc gtcggctgcc gctatggccg cgtttacacc 19980 cgacctgacc accgcgaccc cgcggctgca gtactttcac atcgccggtc cgagcacccg 20040 ggaatacctg tccgaggacc ttcagcagtt tatcgcggcg actggaagct actttgaact 20100 gaaaaacaag ttcaggcaga cggtagtcgc gcccactcgc aatgtcacca ccgaaaaggc 20160 tcagaggctt cagatcaggt tttacccgac gcagaccgac gacacgccca acagttaccg 20220 cgtgcggtac agtttaaacg tgggcgacag ttgggttctg gacatgggag ccacctactt 20280 cgacatcaag ggcgttctag acagaggacc ttcttttaaa ccatacggag gaaccgcata 20340 caatcccctc gcgccccgcg aagccttttt caacaattgg cttgagtcag agccgaacaa 20400 gaccgtcatt acggggttaa tgaccacccc ctaccgaaac gaccagggga acaagaccaa 20460 tactgctgac gtagttaata gcgtgtcggg agtttatccg aaccccaatc tcggaccctg 20520 catcagcgag atgggagcac tggatgatgc aactgctgac cttgtcggct ttgcgggacg 20580 gtttgcgaaa gttacaaacg agaatacgcg tttggcatac ggagcgtatg tgaaaccgtt 20640 gaaaaacgat ggttcgcagt cattaaaccc cactccttac tgggtaatgg acaagactga 20700 cgccaagtat ctgggcgtga tggccgtgga agaattcagc accagtctca cctatccgga 20760 cagcttgtta atccctccgc cctcggacta ctcaaccgtt aacacgggcg caatgaaagc 20820 aaaccgtcct aactacatcg gtttcagaga taacttcatt aatttgctat atcatgatac 20880 gggcgtgtgc tcaggtactc tcaattccga acgctccgga atgaacgttg tcgtagaact 20940 tcaggacaga aataccgagc taagctatca atacatgtta gctgacatga tgtcgcggca 21000 tcactactac gctctgtgga accaggcggt agatcagtac gaccacgacg ttagggtgtt 21060 caataacgat ggctacgaag aaggagtacc aacgtacgcg ttctctcccg agggcacggg 21120 atatggcgct aatcctaatt ctaacctgtc gtttgctgac gtgaaagtgc atcaaaacgg 21180 ggcgggcgac aaaggaaacg agattaacac gggaggtgca acttactcta catacttaac 21240 cgtgggaagt gtgccatctt atgaaatcga tcttgctgct tctcagcggc gaaacttcat 21300 agtgtccaat attgccgatt acctgcccga taagtacaag tacaatattg atggttttaa 21360 tcccgagacg ggtaacgtag accctactac atacgaatac atgaaccgca gggttccact 21420 taccaatgta gtagatctat ttacaactat cggcgccaga tggtctatag accagatgga 21480 caacgtgaac ccattcaatc accatagaaa ctggggtctg aagtataggt cgcaattgct 21540 cggtaacagt agatattgca gattccacat tcaggttcca cagaaatatt ttgctattaa 21600 gaacctgttg cttctccccg gaacttacac ctatgaatgg gtgctgagaa aggaccccaa 21660 tatgattcta cagtccagct tggggaacga tctgagagcc gacggagcct ctattgtata 21720 ctctgaggtg aacctaatgg cgaactttat gcctatggat cacaatacta gtaaccagtt 21780 ggaattgatg atgagaaacg ctaccaacga tcaaacgttc gcagactacc tgggagctaa 21840 gaatgcgctt taccaggttc cagccggctc cactgcactt actattaata ttccagcgcg 21900 tacatgggag gggatgaggg gttggtcttt tacacgtgtc aaagcctctg agacacctca 21960 aatcggtgct caatatgata tcaacttcaa gtactccggt tctattccgt actcggatgg 22020 taccttttat ctcacgcaca cgttcaggaa catgagtgta ttgttcgata cgtctatcaa 22080

ctggcccggc aatgatcggc ttctggcacc caatctattc gaaatcaaac gtaacgtcag 22140 cataaatgcc gaaggattca ccatgtccca gtgcaatatc actaaggact ggtatcttat 22200 ccagatggcg accaactaca attatgtatt taacggctac aggttctggc ccgacaggca 22260 atatttccac tacgattttc tgcgaaactt tgaccccatg acctgtcagg gacctaactt 22320 ccaagatcaa actttatttg atctcatgag ttacgatcca gtgcagactc ccggagccgt 22380 tcagaccaac caagacgcta tccgtaacaa ttccggttac acggctcctc gcagctggcc 22440 ggtatacgga gcacaacagg gagaatcgtg gcccgcgaat tggccgtatc cgcttatcgg 22500 tgacagctca ctcaatccta cccagatagt aaattacaaa aagtttttgt gtgataatta 22560 cctgtggacc gttccgttca gctcggactt catgtacatg ggtgaattga ccgacttagg 22620 gcaaaatccc atgtatacta acaactctca cagcatggtc ataaatttcg aactggatcc 22680 tatggacgaa aatacttatg tatatatgct ttatggtgtg ttcgatatgg taagagttaa 22740 tcaacccgaa cggaacgtgc tcgccatggc ttacttccgt acgcctttcg ctacaggcaa 22800 cgcggtgtaa aaaaactgat taaaaaatgt ccgggaccac ggaaagccag ctgaatcaac 22860 tagtcggggc catgcacctc cgccatcgat tcctgggcgt cttcgacaaa acttttcccg 22920 ggttcctcga tcccaaccgc cctgcctcgg ccatcgtgaa taccgggtct agagcgaccg 22980 gaggtatgca ctggatagct ttcgcgttcg atccgatcgc gcgaaaatgc tacatgttcg 23040 atcccttcgg ttggtcggat cgggaactgt ggaatttgta caaagtgaaa tatgacgctt 23100 ttctcagaag aactggatta agacaacctg ataaatgttt tgaactcgta cgttccgtcg 23160 aagcggttca atgtccgtgt tcggcggcgt gcggactttt tagcgctctt tttattgctt 23220 cgtttgatcg ctaccacacc cgtccgatgg atggtaatcc catcatcgat acggtcgtcg 23280 gcgtaaaaca ctccgacatg tacaaacccg aattccagag cattttgcac cggaaccagg 23340 agcgaatgta tttctggttt atgaaaaaca actctttttt tcgcgcacac gaatcggaac 23400 tgaaacgcga aacggccatc aacagcgttc ccgaaaacca ttaagacatt acactcttgt 23460 aaatttctat atctgtattt tttattttcg cattcgcgac aataaactac atttattttc 23520 gagaccgcgt gcgattattg ctctcgtccg cgcccgttat gcgaacgggt cgcctgtttc 23580 aacgggttta accggagcga tgacctcggt tttaaacccg aaacgctcgt gccaacggaa 23640 ttcgggaacg tgagtcggcg cttgaacgcc catggcgttc aacactaatt ccgtggcaaa 23700 aacgtaagcg tggcgtagat ccatcgcgga aagccgccaa gcgcaggttt tatcggaggt 23760 cttgcgaccg gagcgagagg tgggaccatc cgtgcccgag ggagattgcg gattgcagca 23820 cgtgtacacc atggtatgcg ggtacttctt gtgcgccttc atatcgggac gcgagcgagc 23880 ggtatcggaa gcgatatcgt cggtgccgct cagcttgtat gggatcatgc gacaaacctg 23940 acgcccgctg atgggacctt gaacggcgta attacaatta cagttggtcg atatcagaat 24000 atgctcttcg gcacggcgtt tttccgcatt tggatagaga gccttggtcc actctaagtc 24060 gtgtctcatg gcgctcaccg ctttcgcggc atccgagaag gccatgccgc agcttccaat 24120 cacgtgaggg tacgggaaac cagagtggtc ggcgtccctg gagcacacgg cgttacggtc 24180 gaaccgcaag acgaccactt gtttaccgaa gcggttcttc tcgatttgac cgttttgctc 24240 cgcaatggct ctctttcccg cttcgctagt agggttgagc tcgacggtgc gcggcttcaa 24300 gaacatggca ttaccatgca agcatttggg ggtggtgttg gtggtggctt cccatccgtg 24360 tcgccatacg tgcgctcctt ccggaacgaa cttcggttcg agctccgcca ttccgtacac 24420 catggccgcc agaaaacggc caacttggtt gtggaaggag tcgaaactcg agaaggtaag 24480 tctaaactca ggatgcctct tgcgcacgaa tgtacctcca atcctggtcc aaatggcatc 24540 gtcggggcga atagtggaac cctgccaccg gacatcgagt cggtcgcaaa tcgttgtcag 24600 aatcttcatg gccttctgcg caccgtactc tacgggatcg cttaacggcc tttctaattc 24660 ttcatccgat tcaacgtcga gttcggattt cttttcgtcg acttcttccc gttcctgcgt 24720 ttccttgcgg gcttttttgc tgggcgcggc accccgagac ttagcgctcc tcttcttttt 24780 cgagtccacc atcgctggtt cagcgttctc atgatcgtcg ctagacgcca tctttttgga 24840 ggtagtttta ggcgcggaat cctttaccat gccgctacga tcggtttctc cttcttcctc 24900 ttcttcgtcc tcacccggtg agctctcttc agaactggag aatacttcct cgacttccag 24960 cctcctcttt tgtaccttga cgggatgacc tattggttga acgaaaccgt tttacgacga 25020 cgtcacagac ccctcccata gcccccagcg cggtataaaa gtagatttcc gggtcggtac 25080 gaccactctt atgccatcga gggaggctgc aggagcccgt acctcggcga tccgacggtg 25140 gaagtgtctc cttcggtagc ggtgcttcct gcagcggcct cgtctttggt tggctccaat 25200 ccgattccga gtcgtccgaa ttgggcatcg gagtcttttc cctggctccg aatacctgtc 25260 gaatctcctc gtcgtcttcg tccgagctca ctacccaaag atcgcggtct gctgcctccg 25320 ccatcacggt tcttcctccg gctcctccta gagagttcag atcgagggaa ggagagaaaa 25380 cggcatcatg caaacggctc gcgaagggaa ggccacggtc attctgaatc tcgtcatcaa 25440 taccgcgaag gaaaaaatcg gttggcagcg cttcgtggac gcgatccaac ggtacgtggc 25500 cgaagcttat ggtgctttct tggcgctcga cccagaggcg gcaccaccta gacgcgaatc 25560 cgaccacgcg gtgcgagttc ttatcgatgc tttaggggag gagagagcag ttttagctgc 25620 ttatcgcgtg gcggaggagt tattagaaga taaggagatg ccaaaagaaa gccaagacaa 25680 agaacctttg tctgaaaatg aggaacttgc gcattctttc gcggaaagta ccgtttctcc 25740 gcattctttt gcggaaagta ccgcgacggg tgatataaac gtacccgatg acgagagttc 25800 agtcgcctca caaacttact cgggagagga acgggacgag tctgagtgcg actcggacga 25860 agaagaaaac gacaaggagg cgaacgaagt cgaagcagac gaggcagaag agcaggagga 25920 ggaggaggag gaggaggagg aggacgaaga agatgactct agcccctcca ccgtgctgga 25980 aaacgagcga atacaagaaa acggggaacc cgaccacaat cccaatggag acgagggagg 26040 agctgattct gatgcggata gcggatacta ttcggcggac gcgggacgat tgggatcgtt 26100 actttcggac tcagaacacg aaaacgacac cactgaacgg caagtggaag tgggtatcga 26160 ggacgagtct cctactggtc accatccact tgggactcct gattccgatc gcgatcatca 26220 ggagcaagaa acttctatag agacgaaaca tgtagatccc gtttctactt ttaaaaagtg 26280 cttcgaacgc caagctgctc tgcttaccgg cgcccttaag attccttgcc tgctgaggac 26340 gccgacgaac ccgtctccgt atccgcttgt gcagtaccag ctagagagat tcgtattcaa 26400 tcccgaccca agggtccctc ccgaacaccg cgaagtgcgc tacaatttct accctccttt 26460 catgcgtcca aaagctatcg ctaattatca cattttcgcc gttaccgccc ccatccctgc 26520 tagctgcaaa gcaaacagaa gcgggagcaa attactagag tcctgccgac aaacggcaac 26580 gtttaagcgc ttacctcggt ggcggatcaa cgttcaattc gatgacgggc tcggagaaga 26640 ggtgattcct gtaacagagc tatcagatgc aaagttagtt cctttgcaag aagacatttc 26700 gagattgcag tgggccaaaa tgagaggtga acatattcgg ttctttagct atccctcttt 26760 acacatgcct cctaaaattt cacgcatgct catggaaacc ttgctacaac catttgccga 26820 tgaaaacgat aaaggcgaag gaccaaaacc gtgcgtgagt gatgaggaac tgcgttatat 26880 cgtagatccg gacggcaaaa tgcgcgggga agaactctat aaagctatgc agcggagacg 26940 aaccattgtt accatggcag ttcgttttac cgcgcttctg gaactcatgg aacgcgtcct 27000 cagggagcct tcttcggtca aaaaagcaca agaggtgctc catcatactt tacaccacgg 27060 atttgtcgct caagtacgcg aaacggctaa agttaatctg agtaattacg cgacgtacca 27120 cggaatcacc tataacgacc ctctaaacaa ctgcactgtc gccaagctct ttgaaggtcg 27180 ggacaaggaa gattatgtgc tcgacagcat ttacttattt ttagtcttga attggcaaac 27240 cgcgatgggc atgtggcaac aagccataga cgacaccaca attcaaattt acaccgaagc 27300 ttttacgcgc caacggagag ccatttacgc gctgactagc gtcaccgagg tgtccaaagc 27360 catagtcgat cttctcatgg acggggaccg gttaaccgaa gaaatgcgca aagctctgcc 27420 taactttatt acgcagagcc agttgtctga ctttagacac ttcctaacgg aaagatctaa 27480 cgttcctacg atggcagcac ccttctatcc ctctgacttt attccgctag ccttccgcca 27540 aagcgcacct ctcttgtggg atcacgtcta tctgcttcaa gcggcttact tcctcatgaa 27600 tcacggcgga tacctatggg aaccctccga gagcgatgcg gaatctccgc aatttagagc 27660 ttactgtccc tgcaatctgt gcagccctca tcgaatgtta gcagacaacg tggctctgca 27720 caacgaagtc ctcgccatcg gtacctttga gatcagaagc gcggatggta aaacttttaa 27780 acttaccccg gaactatggg caaatgccta cctagataaa ttcgtggccg atgacttcca 27840 cccgtttacc gtttttcact attccgaaaa ccgctcatct tttggcaaaa actataccgc 27900 ttgcgtcacg gagagccccg aaatcctatc gctgattcgc cagatacagg cctccaggga 27960 ggagtttctg ctcacccggg gcaaaggagt ctacaaagac ccacaaaccg gggaaacgct 28020 agccacttcg gcggcgagcg tcggcgaggc tcgacctgga gctgccaggg ggtgcgtctt 28080 taccgtccgc tcccaccggt gccagcggag gaactccagc gccgcaaaag cctcctaggg 28140 ctatacgggc tgccgatcag atcatctccg gagactcgcg acataccagg cgcgactacg 28200 gggaaccgtc tttatcaatt ccatccgact ggcgatatgg ctcagagaat gctcgacgag 28260 aagaccagag taaccgaaga gccacccccg ccgtcggaag agaccgacgc tcccttccat 28320 acagacgggc tgtacgcggc acgggaagcg atcgagtccg acggcgaaac tctaggaagc 28380 caggatacgg aggaggacct gagtaccatc tcggaggagg aggaagagga gacttacgag 28440 gaagcggaga aggagaatat acctccgtcg aggggggcat ctcgaaaacg ccgctccgca 28500 tcctccctag agaagactct cccgagacct ccgctgagaa agaaatcccg caataacgct 28560 gacgagggta atgcccaagg gcttagaaaa cgtcgcccta gaggtaacta tcgcagctgg 28620 gtcagacacc gggtggctat ctgtcaagct ctacgcgatg cggtattcga ccgcaaaatg 28680 gctgctgaaa ttctcaaaag agtgcacggg ttgttcgtgc ctcctactgt cttaggctat 28740 tacgctcgaa aactcttaga tcttaccgac gaagactctg atcgtcggtc tccctttttt 28800 atgttcttag tgtccgacga tgccgtaatc gacgaagagc cggttccacc gcctctacct 28860 cgtaaacggc gccgtagggc gaataaggag gaaccgaacg cctctgaaac acagcttccc 28920 gaacccgtat ccccagcagt atctgatttg aaagccgaaa tattgaatct gctcgttgaa 28980 atcgaaagtt tcgtccgtaa aaacccgtcg cgccgcgtct cgattcgcaa ccgtacccgc 29040 gagagcatta cgcgccagct gcactacatg aaaagtgaac aaaaactcac caagcttaag 29100 acagatgcgg aaaaaatcct acacctgtgg agatcccttt cataatcccg cttcttttat 29160 agccctccgc tgcgcttcaa catcatgaac ctgctcgacg ccacgcctac cgagtacgtg 29220 tggaaataca acccgctctc aggaattccc gccggcgcgc aacagaatta cggagcgacc 29280 ataaactggg tggtgcccgg aggcaatagt ttcgcctacg cagcggatga gataaggcga 29340 aacaccctga gccccgccgt cacgcgagcg ataaccgcgc gcttcgaagc tgaatcagac 29400 cagcaaccct tcgccaaccc ccgggaaacc gcctacatca ccgctaacgt actggactct 29460 ggctttccaa agtccgcggt ttatcccgta gatccttccg gagtacaaaa agttcagctc 29520 gctggaggcg ccgaggggcg tatgcagctc tcaggcggcc tcactgaagg tcgagtgcaa 29580 ctttcgggtg gtgtattagg acacgtgcct cgccgcagtc ggcgaggcgc cagacctcct 29640

cgatggtgcg gaacagccct cgcgggaaac ggactaccag aagacgccga aatggtttcc 29700 gacacctaca agtatttcct ccgcactcaa ggacccagcc aagtggtctc agaaccgggc 29760 gtatactctc aacggcagtt tatgacaacc tttttacccg ctgttgtccc acatcccttt 29820 gacagtccta atcctaacga ttttcctgct caatatagtg ccatctacaa gggcacaaac 29880 gcttatgagg atgtattttg ggactggtaa gccggtcttc acacttaccc gctgctgtcg 29940 atgctcagtt tcgcaataaa gttcctccaa ttcacgttcg ttaaacggtt cccgtctcgt 30000 cattattacc cgttcgccgc cgtcgctcgc tattcgcgct ctaaaacgtc ttttgagcca 30060 aaagacgtaa ccggggttca ggggttgcgt aaatctaacg atcgcctgat cgttgatttt 30120 caaccaatat tttttaggag catccacttg cgcctccgac atggcgaaat cgactccttt 30180 cgcgttctcc atggggcagc actccagccg aaaacgtccc gcggacagcg aaaacacgca 30240 aaatgcatca aaagtcgcca aaacgcagac ttctgccacg cgcgccggtg tcgacggaaa 30300 tgacgaccta aacctggtgt accccttttg gctccaaaac agcacttcgg ggggcggagg 30360 aggaggaagc ggcggaaacc cctccctaaa cccccctttt atcgacccta acggacccct 30420 ctatgttcaa aacagtctcc tttatgtcaa aactactgca cctatcgagg ttgaaaacaa 30480 gtcactcgcc ctagcttatg actcctcact ggctgtggac gctcaaaatc agctgcaagt 30540 gaaggtagat accgagggtc ccatcaggat ttccccagat ggactcgata ttgctgtcga 30600 cccatcgacg ttggaggttg atgacgaatg ggaattggca gttaaattag atccaaacgg 30660 acctcttacc gcttcatcag cgggaatcaa tataaacgta gacgataccc ttcttataga 30720 agatgacgat gctaatcaag caaaggagtt aggcgttcat ctcaatccta acgggcctat 30780 tactgccgac cgagacggat tagacttaga aattgattca caaaccatgg ttgtcaagga 30840 cagcggaacg tccggcggcg tactcggtgt attattaaaa ccgagtggag gactgcagtc 30900 aagcattcaa ggtatcggag tagcggtagc tgatacgtta accatcacga gtaacaccgt 30960 agaggtaaaa actgacccca atgggtcaat aagctactct gctaacggca ttgccgtaaa 31020 acccgatccc tcaggaccgc tcacttcctc tggtaccgga ctatctgtgg ttacagcagc 31080 cgagggttca atccaatctt ccaatgccgg tttagcggtt aaaaccgatc caagcggacc 31140 gattaccagc ggttcgaatg gcttaaacct gtcatataat gcatcagatt ttactgtgtc 31200 acaaggagtg ctaaacatta ttagaaatcc cagcaccctc cccgatgctt atctggagtc 31260 cggaaccaac tatttgaata attttacggc acaagccgag aattctagtg tattcaaatt 31320 taactgtgcc tatttcctac agtcatggta ttccaatggc ctggtaactt cctctctcta 31380 tttaaagata gacagagctc agttttcaaa tatgccgaca ggtcagtccg cggaaaacgc 31440 cagatatttt accttctggg tacctaccta tgagtctctc aacctgtcac gggtttccac 31500 ccccacgatc acgcctaaca ccgttcagtg gggcgcattt tcgccggccc aaaactgctc 31560 aggtaatccc gcctttcagt acaatctcac gcaaccgcca agcatctatt ttgagcccaa 31620 atcgggttcc gtgcaaactt tccaacccgt attgacagga gcttggaata ccgacaccta 31680 caacccagga accgttcaag tctgcatact gcctcaaacc gttgtgggag gccagtcgac 31740 ctttgttaac atgacatgtt ataacttccg gtgtcaaaat cctggaatat tcaaggttgc 31800 tgctagtaac ggcacattca ctatcggacc cattttctac tcctgcccaa ccaatgaatt 31860 aacacgaccc acataataac ttacacgtgt aaatcaataa aaaagaatcg atgtaaaatc 31920 tttattgatt taatgattca ttgtcgcgtc ccattcggca cattgccagt aatacagaca 31980 attgacagcc tccacgaacc acgcctcact aattaaatgc accttctgtt caggaaaatc 32040 cacctgcatt cttcggcagt agtgacaagg gagtgccccc atttgtccga gcgttgctat 32100 ggcttccacg aaaatattaa ctttattggg gggaacgtaa attaccattc ccagaactgc 32160 accagaagcc ggtgaaggac aaaacaccgg ataaaaatca gtaaacatgg ctgcagccca 32220 agccgccaca aaataaactc cctcaagcgt gggatcatct tctaagcggc tccacaaata 32280 aactcccaga cctgtcccgt cgggctgaca acacaccccc atgggaactc ttttcatatc 32340 gaatccgtga caatcacatt tctcagtaat caactcaaaa tcttctaaac acaagaactg 32400 ataggcatgt gccgcgggaa caaatcccac gttgtttacg gcggcgggaa catcttgtgc 32460 ctgttcagca ccatcagcat ctgctagcgc tctctcggcc atctgtcaca cttgaacgtc 32520 ctcgatttcg gcgtacgggt gactgtcctc ggttaacccc tcgctaaaaa cgaccaatgg 32580 aacgggaacg ggaggagtag gatcggatgg tagccgatac aacacggagc gcaatcccgc 32640 ttcccctgaa tacagggggt tactgctaac tgtccaggta ggagatcttc gtggaaagct 32700 cgacagtcta cgcagccacg aatgtcttcg agatggtgtt ctattggacg cagtacgctg 32760 ccttcggtcc cgccttcccg tagcacacca aaaaatcagc aataaactca cggtagttgc 32820 tgtcaaaaag gtgcaacatc caaacacggt aaaaacaaag gtcagttcgg atatacggtg 32880 actcaggaac agatagctga caaccacgga agcgcagtaa gaacatacta ctgctgaaat 32940 gcagaacagg atacatagtc ttctgatccg agcgtcgctc ggtacctaac aataaaagaa 33000 aggcgatttt ctatcacctt ctactctagc acgtcttttt gacatgcaat tgcgtaagta 33060 actgagttgt gcaatataag ctagtggcaa cggttgcttg aggggacacc ccgggcatgc 33120 gctctcagga tgatgatcgc aataagtacc atgcacatgc aatacccatt cctccacctc 33180 tctataggta gcccgtctaa aggcgggaat gtaacacaac cccttagaaa caataggcca 33240 aacagaaggg tcggccatct ccctcaacat tgatcgtgtc attaacggtt gagtcacggg 33300 actaccactg ccgctcgttt ttaaaataac agtagctaca acattggtac ggcatgcgcc 33360 cagaaaaagg acaggatagg gcaagaacgg taactccctt ggcacctttg tcgtagacac 33420 ctcgtaacta acagtttgta acgcgaactt aaaagctaaa ctgttatcta acagaatccc 33480 tgatcgcaaa aatacaaaat cggacttgtg tctcctgata atccacttgc gatgcctatc 33540 taccaactcg gtcttggcag caaaaagcct tctcgaaaga acccgtaaaa actcctcctt 33600 ctgatcggga tccagttgca atcttgcttg aacccttaca atcacgttca agatgatcct 33660 cggccccgac tttaccctca catactttgc acatctacaa ctgactacgg aatacaggga 33720 ctcgtgccac agataatgtt gtggcttttc gaagaggtcc atgctcatga ttgtccgatt 33780 cgtgtgaaat tccatcatcc gttctaggga ctcctgagtc gcaccctgtc tcatcagtag 33840 actaaaccat atcactccgc actcattttg aaatccgcag cccctggtaa gggctaccac 33900 ctcctgtaga attacaagga tgcaagatga aatcagaata tagagaagaa tcagcgagcg 33960 catgcagcac atgcataagc acttccatga aactcaccga ctcagtacac agaggcatga 34020 tgtcgagact gatgattttt tattgatcaa gtaagtccac ctgaaacata ttaaacaaca 34080 tcataataaa tatcgcggta acatttacaa cggtgatggc agaatatata catcacgaca 34140 catcccgtga aacacgcagc ggctacaccg cccgtcacaa ccgcgatacg cgtaactcta 34200 tcttcccaaa attcacggat cagagtgtaa aactctcctc cgagatgagc aatggtgggg 34260 tgcccttcgg ttttctgctc cgagtacgta acagcgtcac catcatggct acaaatgcaa 34320 gtcctacgac aattaaaaaa agagcttccg tcgaaacacc cgtcgaacct tcttccgatg 34380 gaatctggac tggtgcttga agtgcctcca tccgaggaaa gggttccgat gcgtttagag 34440 ttatcattat cgcgtctgcg ttcgtagccg tctcgttcca accatattcg gaagtcggag 34500 taggatctga tgactcccaa accgaggtag cgttcaccga agtatagttc gcaataggct 34560 ccgctgtcgt aaccttctcg acaacacggt ttaccggtga tgaaaccgtt actttcaatt 34620 ttataacttc gaattgaagg aagaattcca gatcataaac atcttcagca aagaaagtca 34680 cgttaaactc tttcttcatt cgattcagat ggaaaacaca ctgagaactc gttacacacg 34740 tatcccaata actcactagt ctatgagtcc ccaccctgtc agtcagatga acactcatca 34800 accaaaacgg tgacctactc aaatctatct gtaacttgtg tccgaccgtt acattcatat 34860 caggtaattt actgacaatc ataccggcgg ttgtattctt atcgaccaag catcctgccg 34920 tcggcggaac gtttatatgg gcacctccat tggggtttaa cattgtacgg tattgcataa 34980 tcggaaaatg ctccttagta tacatacaat agtgagacgg gcgtccgaga ggatccggtg 35040 tgcgtgtaca atacatagtg cgaacagaac ccccggtcat aatttgtcta tttgctaagg 35100 gagcaacaga atcggatacc tggtaaaatc cctgaggcgt aatgacgtga actaaatata 35160 tacgggcatt cggatttaag tgatggatcc acagagaacc ttccgccaac tcggtatacg 35220 ctatctcggt ggttacatcg tatttcccat atcttaaagt ctgcacacta tagcctccca 35280 tatatacgct gctatcatgt gaagcaacta ctataatcat aatagaagaa acaaagtcct 35340 tcttaaaatt tggggaatac gacatttccg tcatatactt tatgctgtaa tggttgaagt 35400 aactgtcata tctataatca cgactaagtg tatacccatt ccaagtactc cttaccggtc 35460 ctaccctcga cccttcctgt aatacatgga acacaggcaa acctgtcttt acatccaggg 35520 ttttcaaaaa ctttggaaca atcattcgat gatagcatcc acttaaacta aacgtattac 35580 cccaggaaaa ttcctcacat actgtcgctc cccaaacatt ttctccgcat actcttccgg 35640 accatccacc gtgtatacca cagcttctac cgcccttatc ttccaccatt aaatcttccg 35700 aagccgtgta cccaccagtg atccacgaat actgagtctt tagaacgacc acataatccg 35760 catccaactt atctaatcca ccgcttacaa atgtcggttc tagcccgact attctgatac 35820 accgatggcc cttgctcatt tgtgaatact gtctacatat cgtcgagcaa gcatgagctc 35880 ctgccgagtg acctatacag tgcaattttt tagcgtcggg gatatctacc agaaacttcc 35940 taaaatccgc atgtaaacca tccgtatcat agtacaaccg atagtccacc aacacaacac 36000 ctacggcggg agtcatcttc tgatgaaatc gcaacacctt cctaaaatcg tcatatccca 36060 gataccatcc tggtataagc agcactatgt ctgatttccc ggcgaaggca ttgtgcattt 36120 tatggtatac accatactcg tatctcgcat cgtggaactt cggtacacca ccatgcgatc 36180 cataccacgt gattctcgaa ggaagtctgt tcggatttcc ggcgcttttc ggagcttcca 36240 gtttagtagc cggaggcatt gccacacagg accgattgac caaatcatgg caaccttctg 36300 atttcgtacc cgaaaatgcg gaggccccga ggagcacccc gaataactga aacagaacgc 36360 aaaatttaga ccggtaccct cccatacgat tatcacgtcc caaagccaaa tatcacggta 36420 ctcaccagcg caaaaatctt catgaagatc tggggcaaga gccgcacaac agtccacctg 36480 aatctgagac agcttaccgg cgactgtacc aaaccggtgt gccaccgccg aatttattct 36540 ctcaatctat taatgtttaa ctaaacaaac ttgctgacac ttgataaata tttactgacc 36600 tagacaattc cgagaacttc cttattaccg tgactatgca gtatcaagat aaacccttcc 36660 aaagttcgac tgagtacacg gtgtacttcc tgtcataaaa tatcacttct aaatatagtc 36720 ctaggtagaa actataaatg gtaagagaca acacactgat tcttagaaaa acctcacaga 36780 acaagatgaa cctcacatta tcctgactct tccttacgaa ctgaaataac cggtaagtca 36840 taaaaatgat tttccttttg acattgcgta cgcggaagca aactagaaaa atcgaacttg 36900 gaattttcca agtcgagtta actattaact tgatgacgtc aattggatta atcattaact 36960 cgaattagtt aatgattaac tagatctggt taatgattaa ctagtgacgt cactagttaa 37020 tcattaagct ttctatgcat atgcatgaga gattcatgca tatgaatgag agtctcatcc 37080 atattcataa acctcatcca tattcatgag cctctcatgc atatgcatgt ttttacatgc 37140

atatgcatgg acctcattca tattcatcaa ttcgttaatg tataacgcta tgactcactg 37200 tatatgttta cgttgcctag caacgttaat gatttacctg ctgacgtggc agctccgcct 37260 cccggtaaat catttacctg gactttgttc tttatgttta ttcaccatgg caacgctacc 37320 atatatggac atccgactcc gcctccccag ttatacatta acgatggcgt gataggcgga 37380 gctctctccc attggctctt aatgacgtag tccgggttaa ccataagcca gaaccgccta 37440 tataggtaga gcaggtagac ccggaacacc attcccatac ggacctccat agagtgcgga 37500 cctctacggg ctctccatac cggtaaatat tttattccat ttaatccaat caaataaatc 37560 aataatcaac tcaatgctgt gattctgcct caaattcaat ggtgattttc tttaataaaa 37620 agcccacccc ccttggcacc cccctgtaca cccccctgta caggcgacca ccccctatgg 37680 acacccccct gtacaggcga ccacccccta tggacacccc cctgtacagg cgaccacccc 37740 ctatggacac ccccctgtac aggcgaccac cccctatgga cacccccctg tacaggcgac 37800 caccccctat ggacaccccc ctgtacaccc ccctgtacat tttctcccat aggctacaat 37860 ggaacactgc cccctagtgt ctcccgctcc atgggacccc tatgaggtgg gcaccatctc 37920 atttgccgca atggagctca tccacgaggg ggcgccattg aagctagggg accgcataga 37980 gagcctggcc aatggggtgc tttggaatcc ggatatcccc gcccaactct tcaactgcat 38040 ttctattcgc tcatggggat cacatgggaa gcgcgtcata ttcaaaggcc acacccaccg 38100 gatgttccac gcccagctta ggatccgtag caccgccccc gttactagga aacaggccgg 38160 aagcctactc ctcagcctat cacagaggct cctgtgtttc cccgcccgct ataataccca 38220 ccccctcgtg atgcaattgg gggtggagtc taatgccatg ggacttccta tatattccaa 38280 gagggccctc gagatggcgc tcaagagcat gcgggtgcgc attgcgccta acagccacca 38340 ggtggcgcca cctgaaatag gaaagacctg tacggtgaag cccctcaaag aagtggagac 38400 cctccagcag ggggtcttca gtaccaccga tctaaaaaga gcacttccag aatggacttt 38460 tcgccgactt tttaacgaaa ttccctatat ttgtggctgg aagattggca ctgctccaaa 38520 gggcggagaa atggatggtt acgctccacc ccgaatccag agccccgccc accgaaggac 38580 ggaaggaccc gcccactctc caagacctcg cccggctggg cgtggtcgaa aactgcctca 38640 agatgaggag gcggggcata agccctaggc accaccccta ctttcaataa accaatcaga 38700 acagagcact tgactcctcc tatttgtggg cggggctatg ggtacggtat agagcgcccc 38760 ctggcggccg ttggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 38820 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 38880 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 38940 tggcgcgggc cggtcaatca ttaacccggc gcccagcacg ggcgccccct gccggccgga 39000 gacagacact gcgtgtctgc ggagctcccc ctggcggccg agggccgcca ctgcacccgc 39060 gccggactta gtctctggcg cgggccggtc aatcattaac ccggcgccca gcacgggcgc 39120 cccctgccgg ccggagacag acactgcgtg tctgcggagc tccccctggc ggccgagggc 39180 cgccactgca cccgcgccgg acttagtctc tggcgcgggc cggtcaatca ttaacccggc 39240 gcccagcacg ggcgccccct gccggccgga gacagacact gcgtgtctgc ggagctcccc 39300 ctggcggccg agggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 39360 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 39420 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 39480 tggcgcgggc cggtcaatca ttaacccggc gcccagcacg ggcgccccct gccggccgga 39540 gacagacact gcgtgtctgc ggagctcccc ctggcggccg agggccgcca ctgcacccgc 39600 gccggactta gtctctggcg cgggccggtc aatcattaac ccggcgccca gcacgggcgc 39660 cccctgccgg ccggagacag acactgcgtg tctgcggagc tccccctggc ggccgagggc 39720 cgccactgca cccgcgccgg acttagtctc tggcgcgggc cggtcaatca ttaacccggc 39780 gcccagcacg ggcgccccct gccggccgga gacagacact gcgtgtctgc ggagctcccc 39840 ctggcggccg agggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 39900 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 39960 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 40020 tggcgcgggc cggtcaatca ttaacccggc gcccagcacg ggcgccccct gccggccgga 40080 gacagacact gcgtgtctgc ggagctcccc ctggcggccg agggccgcca ctgcacccgc 40140 gccggactta gtctctggcg cgggccggtc aatcattaac ccggcgccca gcacgggcgc 40200 cccctgccgg ccggagacag acactgcgtg tctgcggagc tccccctggc ggccgagggc 40260 cgccactgca cccgcgccgg acttagtctc tggcgcgggc cggtcaatca ttaacccggc 40320 gcccagcacg ggcgccccct gccggccgga gacagacact gcgtgtctgc ggagctcccc 40380 ctggcggccg agggccgcca ctgcacccgc gccggactta gtctctggcg cgggccggtc 40440 aatcattaac ccggcgccca gcacgggcgc cccctgccgg ccggagacag acactgcgtg 40500 tctgcggagc tccccctggc ggccgagggc cgccactgca cccgcgccgg acttagtctc 40560 tgacaccgca tttactttca acaccttttt ttatttcagc acaccgaaat ctcgtccgtg 40620 taaaccttgt ccccaaacct cagtcccgta catcgaaccc atccacatat atctcgcaca 40680 gctccagtgc ttacaagata tttccctact ggataattag gatcttgaca ccacttattc 40740 aaatacatcc ggaaccactg tccgtctgga acttggcttc tgtagaactc ctcacgaaag 40800 tagcgcttca ttgctctaga acacttatac aaatgtttat ctctcaggtc gaacatcgtg 40860 aggcaaatca acagcgtcaa atgacaagca caagaaccgc ccggctgaag cccgacgcag 40920 taacccatag gatgtccgac accgatgtta gcttcttcgt ggtacggata ccacagctcc 40980 ttgatgtctt gaactagctt ccagaagatg gcatcgtgca tacaccacgg aggtacggca 41040 acaccaaaca ccacgtacaa aggcatctcc cgacactctg caattacaac tagcatcaga 41100 aatacatcca gccgctattt gaaaaaaaca acaggatctc acccaaaaag aaaaagtact 41160 taccccgaaa ccaccaggaa atccccctta cgtgataatc cggatctcgc tgacgctcaa 41220 agaagccgcc ttaaataccg ctccttatct ctcttcgatt acctcagtta cgcccatttc 41280 ttatcaggta agatacgcag gtaagatacg cccttccaca tcagcacgga atgtgctgac 41340 cagcttaccg cgcccgtttt taatatccgt taattagtaa cttaggtgaa tcaccaaacc 41400 acctgccggc atatatctac acccttgata cggcggcatc cattgaagtc gagcgccacc 41460 atggagtcaa ctaccgtatc ctcgattctg ctcctatcct tcttcgtatc ttccatcgaa 41520 tcctatccgc tcctgcataa cttcacggcg ctcacgggat ccgtgcttac tcttccctac 41580 aaaggacatg accgaccctt taaattcgaa tggcggctga cagatcagac caaagtagca 41640 atctcggatc ctagcaccgg catcaactac ccgtcgggtc cactcaaagg aagagtacat 41700 ctaaacggca gcgccctcat cgttacatcc ctacgactta gtgacgccgg tacctataca 41760 atcctctcag aggacaacac aggaactgaa atcggatact cctattacgt cgaagtcaga 41820 ggtatagaac tttcccctcc tttttctaca tgatatcccg ttccctcttt tttcttaccc 41880 ctagcttatt cttcctagaa ccgatgcaag ctcccaccct ctatacggat cgctacaatg 41940 actcatctgc catacgcctc acatgccaag cttccaacaa tctccaccgt aatatcacat 42000 accactggaa aacagattta atacagccaa ctaactcgac tcgatctatc accttaaaca 42060 tcgaagacta tatatccgtt acctgcacag tcactgatgg agtcagcaaa aacagcatca 42120 cgattatggt tccgttgaga ggtagtattc ggtttcccgt tatctcccta tactacgtaa 42180 tcatgcccac ccttttaacc cgtatcttta caacagatcc ttctccacct acgccttacg 42240 gtttgcatcc cactatcatt ttcctgagtg tcctctgcat gatcctcatc accgcaatta 42300 ccgtctactt tgtgaagaag cattgctgcg ataagcaata taaaataact tgcattaacc 42360 cttaccgaga atgcttcggt ggcgccggtc tcgtttaatc tcgtttaaat aaaaaccact 42420 ataatccccg cttcctataa tctctacttc ctatgattaa atgggtaatt aaatgatggg 42480 taattaagtg gtgtcaatga tcaaataatg attcggaaac gatttcggga aaaatgattt 42540 taggaatcaa tgatcattat caaaaataaa tgatttattt ataagacgtg tttttgatga 42600 ttgatacgat tagtaaatca catgattaga aacacatgat tagaaacaca gtttaatcat 42660 caatcgaaat caccgcgcaa gatatggaca gtcacgttct cctctgatgt cctatccata 42720 gattcgctgt ccgtagaatc agaggcgcag agcatccgcc gaaaccagaa agttatcgct 42780 accatcagca ataacaacat aattaacgcg atcggcagcc acactggaaa ctcgccggtc 42840 tgattggata ccagtcctcc gccggtctta taagtggtct cctcgatctt ctgatcctcc 42900 tgagtcaaac caggaattac cactacgtaa aatccggtct ttgaatcctg ttgtaagtct 42960 agcgtattag tatacagtcc cgaatcggca ggtgttacgt ctctaatagt aacgcaattg 43020 gatgtggcat tgtaatgcgt gcggctctta tagttagcat acactatggg gttacccaga 43080 gctttataaa gctgtaatac gaacacggta gacgcgtgct cgttagagga ctgaaactcc 43140 cactccgtta tagaagcgga gctagcatcc ttcccacaca tgcgtaactc tccaccttcc 43200 aaaactaata tacggctcat gttatgcacc tcaaccgtcg cattgcccgt aatatccttc 43260 acggtatact gtatggcata cggtttccca ccgacaaagg aaccgtaagt caaccagcac 43320 ccgttattag ccttcagctt actcaggtaa atagtattgc aaccactatc taccctcgtt 43380 ctacccatgt acccgctttc aagtccaggt tctttaaccc tcggaatcac aaatgccgcc 43440 ctctccatct tattgcaatt gggggacggg gaatgatacc agaggatcat agtatggtcc 43500 ctgcactctc ctgtccgtag agccagctcc gggtcccggg gagcgccaaa gcagagtccg 43560 aggacagcag ccagcagcag ggcaacggac ttcatgatga atgcgggaac cagccagcag 43620 cagcaggtag aaggtgcgag tactcgaggc aggcgggaat aacaccgcac gctttccacc 43680 ccgcttaaat acacaaccca caccggtcat ggtcaaacat tacctagcct actcagcacg 43740 gaaaagtact cacccttaac acagttcgac ttatcaggta cacacccgta accttcgcac 43800 cccaaagcgc acagtcagca ccataaggaa gtcgtaaagt taccctttga ttgcattgtc 43860 atcataaaaa acgctgagca aacggaaacg ccccaagtac aaatctctct tccgccacgc 43920 cctcgttaat caataactag tcgccgatac atatataccc cctccgttcc atcacaccta 43980 cactaccttc tctccgcaca ggcaacatct caatccttac tcttccgaat ccacttcgcc 44040 gccttcgaca tgaatccggt aagtatcaac ttttttctaa gcttatgctt ctgacacctc 44100 tgatcttaat accgcacccc ttttttctca aaaaaggttt cgtgcttaac gcttcttcta 44160 tgcgtcatca tcccgagagc cgatccgctt ccaatcaccc ccgcctctaa acccgctacc 44220 gactccgccc ggaccggcgc ctccgtcatc acgacccact tacccgcatc tccgagcgcc 44280 accccgtgcg acgaaatctt gagcgaggac tgttggttcg aaaatgccag cggtgattac 44340 caacccctac cctgggaacc gaaagaggaa acggtttcag atcaaaaccc tctcaccgca 44400 accgacccca tcggtgaccg aatccccgct atcatccagt cccagtcccg cgcttctaac 44460 cgtccgacca gtaaggaaca cacttctact atcgcatcca tctcgaccgt agtcggcatc 44520 gcggtcctcg tcatcctgac cctcgtagcc tactttacca agtaccccaa accgagaccg 44580 cccagatcca tctacatagg agtagctcca cccgatatgg aactcaagga aatataattc 44640 cagccccacc cttaacccta cctttccatg agtcagtatt ttcaataaag ttatattgca 44700

gtattaattc cgttgtttcc gcgttctttc tctcgcgcgg acaaagtccg ttcgaccagg 44760 aagtccggta tacgtcattc cgcggtgtca tgatgacgcg cataactcac gactgccatc 44820 tgccggacaa acgcggtact acacttaaca cataaacacc cgcctttttt cgattcccac 44880 cataatcagg ctttggtaaa aattcgcaga ttctaaaatc cgtattcctg cgcccgcgat 44940 aatagatcac gcccacgaca cgccctagcg ctcttatcac acggtctctg ctgccatcta 45000 gcgggcggga gcggtagtgc gaggtgacag cagcgtcatt catgtaggta tatatagatg 45060 atg 45063 <210> SEQ ID NO 2 <211> LENGTH: 714 <212> TYPE: DNA <213> ORGANISM: Aequorea victoria <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(714) <223> OTHER INFORMATION: Enhanced green fluorescent protein (EGFP) <400> SEQUENCE: 2 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 60 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 120 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 180 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 240 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 300 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 360 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 420 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 480 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 540 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 600 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 660 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caag 714 <210> SEQ ID NO 3 <211> LENGTH: 711 <212> TYPE: DNA <213> ORGANISM: Discosoma sp <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(711) <223> OTHER INFORMATION: mCherry Red protein <400> SEQUENCE: 3 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag 60 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc 120 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc 180 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac 240 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc 300 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac 360 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta 420 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc 480 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct 540 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc 600 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa 660 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta g 711 <210> SEQ ID NO 4 <211> LENGTH: 553 <212> TYPE: DNA <213> ORGANISM: Human cytomegalovirus <220> FEATURE: <221> NAME/KEY: promoter <222> LOCATION: (1)..(553) <223> OTHER INFORMATION: Immeadiate early promoter <400> SEQUENCE: 4 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 60 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 120 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 180 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 240 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 300 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 360 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 420 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 480 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 540 ggtctatata agc 553 <210> SEQ ID NO 5 <211> LENGTH: 584 <212> TYPE: DNA <213> ORGANISM: Cytomegalovirus <220> FEATURE: <221> NAME/KEY: enhancer <222> LOCATION: (1)..(584) <223> OTHER INFORMATION: CMV enhancer/ chicken beta-actin promoter (CAG) <400> SEQUENCE: 5 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 60 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 120 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 180 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 240 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 300 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 360 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 420 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 480 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 540 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcg 584 <210> SEQ ID NO 6 <211> LENGTH: 1190 <212> TYPE: DNA <213> ORGANISM: Homo sapiens <220> FEATURE: <221> NAME/KEY: promoter <222> LOCATION: (1)..(1190) <223> OTHER INFORMATION: Human elongation factor 1 alpha (EF1A) promoter <400> SEQUENCE: 6 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 60 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg 120 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 180 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa 240 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt 300 gaattacttc cacgcccctg gctgcagtac gtgattcttg atcccgagct tcgggttgga 360 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt 420 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt 480 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt 540 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt 600 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg 660 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct 720 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaac aggctggccc 780 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct 840 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa 900 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca 960 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt 1020 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc 1080 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca 1140 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga 1190 <210> SEQ ID NO 7 <211> LENGTH: 114 <212> TYPE: DNA <213> ORGANISM: Fowlpox virus <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(114) <223> OTHER INFORMATION: Promoter Sequence for L2R (Fowlpox promoter) <400> SEQUENCE: 7 catagtaaat atgggtaact tcttaatagc cataattaaa attgaaaaaa aaatatcatt 60 ataaaacgta aacgaacaaa aaacattaat ttaaatttgc caataacaaa ccat 114 <210> SEQ ID NO 8 <211> LENGTH: 276 <212> TYPE: DNA <213> ORGANISM: eukaryotic cells <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(276) <223> OTHER INFORMATION: Promoter Sequence for B-actin (minimal) <400> SEQUENCE: 8 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa 60 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg 120 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg 180 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg 240 cggcggccct ataaaaagcg aagcgcgcgg cgggcg 276 <210> SEQ ID NO 9 <211> LENGTH: 516 <212> TYPE: DNA <213> ORGANISM: Woodchuck hepatitis virus <220> FEATURE: <221> NAME/KEY: misc_feature

<222> LOCATION: (1)..(516) <223> OTHER INFORMATION: Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) <400> SEQUENCE: 9 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 60 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 120 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 180 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 240 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 300 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 360 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc 420 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 480 tccagcggac cttccttccc gcggcctgct gccggc 516 <210> SEQ ID NO 10 <211> LENGTH: 1695 <212> TYPE: DNA <213> ORGANISM: Influenza A virus (A/chicken/Mexico/435/2005 (H5N2)) <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1695) <223> OTHER INFORMATION: Segment 4 Hemagglutinin gene <400> SEQUENCE: 10 atggaaagaa tagtgattgc ctttgcaata atcagcattg ttacaggtga ccaaatctgc 60 attggttatc atgcaaacaa ttcaacaaaa caagttgata caatcatgga aaagaatgtg 120 acggtcacac atgctcagga catactagaa aaagaacaca atggaagact ctgcagtctt 180 aaaggagtaa agcccctcat cctgaaggat tgcagcgtag ctgggtggat tctgggaaat 240 cctatgtgtg gtgaattcct gaatgtgccg gaatggtcat acattgtgga gaaagacaac 300 ccatccaatg gcctgtgtta ccctggaaac ttcgacaatt atgaagaatt gaagcattta 360 atgagcagca caaatcacat cgagagaatt cagatatttc ctaggagctc ttggtccaac 420 cataatgcct catcaggagt gagctcggca tgcccgtcca atggtatatc ttcctttttc 480 cggaatgtag tgtggttgat caagaaggat aatgtgtacc gaacactaaa gaagaactac 540 accaacacca atgtagaaga ccttttaata atgtggggag ttcatcaccc tactgatgca 600 actgagcaga caaaactcta ccagaacctg aacacttatg tatcggtggg aacatcaaca 660 ctaaatcaga ggtcaatccc gaaaatagcc accagacccc aggtgaacgg acagagagga 720 aggatggaat ttttttggac aatattaaag ccgaacgatt caatcttttt tgagagtact 780 gggaacttta tagctcccga atatgcatac aagatcacta aaaaagggaa atcagcagtc 840 atgaaaagtg aactggatta tggcgactgt aataccaaat gccagacccc agtgggtgct 900 gtaaattcca gtctgccttt ccacaatgtc catcccttta ccattgggga atgccccagg 960 tatgtaaaat cgaagaaact ggtccttgca acaggattaa gaaatgtacc ccaaaaagaa 1020 acaagaggct tatttggagc aatagctgga ttcatagaag gggggtggca aggaatggtg 1080 gatggatggt atggatacca tcatagcaat gagcagggta gtgggtatgc tgcagacaaa 1140 gaatctacac agaaagcaat taatggaatc actaataaag tcaactcaat cattggcaaa 1200 atgaacactc agttcgaagc cattgggaaa gaattcaaca acctagaaag gagaatagaa 1260 aatttgaata agaaaatgga agatgacttt ttggatgtat ggacttacaa tgcagaactt 1320 ctagtgctca tggaaaacga gagaactctg gatctccatg attcaaatgt caagaattta 1380 tacgataagg tccgactcca actgagagat aatgcaaaag aactaggcaa cggatgcttt 1440 gaattctacc acaagtgtga caatgaatgc atggaaagtg tgagaaatgg aacgtatgat 1500 tacccacaat actcaaaaga gtcaaaactg aatagagagg aaatagaagg ggtcaaatta 1560 gaatcaacag ggacttatca gatactttca atctattcaa cagtggcgag ttccctagca 1620 ctggcaatca tggtagctgg tctatctttt tggatgtgct ccaatggatc attgcagtgc 1680 agaatttgca tctaa 1695 <210> SEQ ID NO 11 <211> LENGTH: 1723 <212> TYPE: DNA <213> ORGANISM: Influenza A virus (A/chicken/Jalisco/CPA1/2012(H7N3)) <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1722) <223> OTHER INFORMATION: Segment 4 Hemagglutinin gene <400> SEQUENCE: 11 ggatacaaaa tgaacactca aattttggca ctcattgctt gtatgctgat tggagctaaa 60 ggagataaaa tatgtcttgg gcaccatgct gtggcaaatg gaacaaaagt gaacacatta 120 acagagagag gaatcgaagt agtaaatgcc acagaaacgg tggagactgc aaatactaag 180 aaaatatgca ctcaggggaa aagaccaaca gatctgggac aatgcggact tctaggaacc 240 ctaataggac ctccccaatg cgatcaattt ctggaatttg acgctgattt aataattgaa 300 cgaagagaag gaaccgatgt gtgttatccc gggaagttca caaatgaaga atcactgagg 360 caaatccttc gagggtcagg aggaattgat aaagagtcaa tgggtttcac ctatagtgga 420 ataagaacca atggggcgac aagtgcttgc agaagatcag gttcttcctt ctatgcggag 480 atgaagtggt tactgtcgaa ttcagacaat gcggcttttc cccaaatgac taagtcgtac 540 agaaatccca ggaacaaacc agctctgata atttggggag tgcatcattc tggatcggct 600 actgagcaga ccaaactcta tgggagtgga aacaagttga taacagtagg aagctcgaaa 660 taccagcagt cattcacccc aagcccgggg gcacgaccac aggtgaatgg gcaatcagga 720 aggattgatt ttcactggct actccttgac cccaatgaca cagtgacctt cactttcaat 780 ggggcattca tagcccctga cagagcaagt ttctttagag gagagtcaat aggagttcag 840 agtgatgttc ctttggattc tggttgtgag ggggattgct tccacaatgg gggtacgata 900 gtgagttccc tgccattcca gaacatcaac cctagaacag tgggaaaatg ccctcgatat 960 gtcaaacaga caagcctcct tttggctaca gggatgagaa acgtcccaga gaaccccaag 1020 gataggaaga gccgacatcg aaggaccaga ggcctttttg gagcgattgc tggattcata 1080 gagaatggat gggaaggtct cattgatgga tggtatggtt tcagacatca aaatgcacaa 1140 ggagaaggaa ctgcagctga ttacaaaagc actcaatctg caatagatca gatcacaggc 1200 aaattgaatc gtctaattga caaaacaaat cagcagtttg aactgataga caacgaattc 1260 agtgaaatag aacaacaaat tgggaatgtc attaactgga cacgagattc aatgactgag 1320 gtatggtcgt acaatgctga attgctggta gctatggaaa atcagcacac aatagatctt 1380 gcagactcag aaatgaacaa actttatgag cgtgtaagga aacaactgag ggagaatgct 1440 gaagaggatg ggactggatg ctttgagata tttcataagt gtgatgatca gtgcatggag 1500 agcatcagga acaacactta tgaccatact caatacagag cggagtcatt gcagaataga 1560 atacagatag acccagtgaa attgagtagt ggatacaaag acataatctt atggtttagc 1620 ttcggggcat catgttttct tcttctagcc attgcaatgg gattggtttt catttgcata 1680 aagaatggaa acatgcggtg cactatttgt atatagtttg aga 1723 <210> SEQ ID NO 12 <211> LENGTH: 1723 <212> TYPE: DNA <213> ORGANISM: Influenza A virus (A/chicken/Guanajuato/07437-15/ 2015(H7N3)) <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1722) <223> OTHER INFORMATION: Segment 4 Hemagglutinin gene <400> SEQUENCE: 12 ggatacaaaa tgaacactca aattttggca ctcattgctt atatgctaat tggagctaaa 60 ggagacaaaa tatgtcttgg gcaccatgct gtggcaaatg gaacaaaagt gaacacatta 120 acagagaggg gaatcgaagt agtaaatgcc acagaaacgg tggagactgc aaatactaag 180 aaaatatgca ctcaggggaa aagaccaata gatctaagac aatgcggact tctaggaacc 240 ctaataggac ctccccaatg cgatcaattt ctgaaatttg acgctgattt aataattgaa 300 cgaagagaag gaaccgatgt gtgttatccc gggaagttca caaatgaaga atcactgagg 360 caaatacttc gaagatcagg aggaattgat aaagagtcga tggatttcac ctataatgga 420 ataagaacca atgggacgac aagtgcttgc aaaagatcca atccttcctt ctatgcggag 480 atgaagtggt tactgtcgga ttctgacaat gcaactttcc ctcaaatgac tatgtcgtac 540 agaaatccca ggaacaaatc agctctgata atttggggag tgcatcattc tggatcggct 600 actgagcaga ccaaactcta tgggagtgga gacaagttca taacagtagg aagctcgaaa 660 tacctgcagt cattcacccc aagcccgggg ccacgaccaa aggtgaatgg gcaatcagga 720 aggatcgact ttcactggct actccttgat cccaatgaca cagtgacctt cactttcaat 780 ggggcattca tagctcctga cagagcaagc ttctttagag gagagtcaat aggaattcag 840 agtgatgttc ctttggatcc tggttgtaag ggggattgct tccacaatgg gggtacgata 900 gtgagttctc tgccattcca gaacatcaac cctagaacag tgggaaaatg ccctcggtat 960 gtcaaacagc caagcctcct tttggctaca gggatgagaa atgtcccaga gaaccccaag 1020 gataggaaga gccgacatcg aaggaccaga ggcctttttg gagcgattgc tgggttcata 1080 gagaatggat gggaaggtct cattgatgga tggtatggtt tcagacatca aaatgcacaa 1140 ggagaaggaa ctgcagctga ttacaaaagc actcaatctg cgataaatca aatcacaggc 1200 aaattgaatc gtctaattga caaaacaaat cagcagtttg aactgataga caacgaattc 1260 agtgaaatag aacaacagat tgggaatgtc attaactgga cacgcgattc aatgactgag 1320 gtatggtcgt acaatgctga attgctgata gctatggaaa atcagcacac aatagatctt 1380 gcagactcag aaatgaacaa actttatgag cgtgtaagga aacaacttag ggagaatgct 1440 gaagaggatg ggactggatg ctttgaaatg tttcataagt gtgatgatca gtgcatggag 1500 agcatcagga acaacactta tgaccatact caatacagag cggaatcatt gcagaatagg 1560 atacagatag atccagtgaa attgagtagt ggatacaaag acataatctt atggtttagc 1620 ttcggggcat catgttttct tcttctagcc attgcaatgg gattggtttt catttgcata 1680 aagaatggaa acatgcggtg cactatttgt atatagtttg aga 1723 <210> SEQ ID NO 13 <211> LENGTH: 2652 <212> TYPE: DNA <213> ORGANISM: Gallid herpesvirus 1 strain USDA <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(2652) <223> OTHER INFORMATION: Laryngotracheitis virus (ILT) glycoprotein B <400> SEQUENCE: 13

atgcaatcct acatcgccgt gaacattgac atggctagct tgaaaatgct gatctgcgtg 60 tgcgtggcaa tcctgatccc atctacccta tctcaagatt cacacggaat tgctggaata 120 atagaccctc gtgatacagc cagcatggat gttggaaaaa tctctttctc cgaagccatt 180 gggtcggggg caccgaaaga accccagatt agaaacagaa tttttgcgtg ctcatctcca 240 actggcgcca gtgttgcgag gcttgcccag ccacgacatt gtcaccgaca tgccgattcg 300 actaacatga ctgaaggaat tgccgtagtc ttcaagcaaa acattgcccc gtacgtcttt 360 aatgtgactc tatactataa acatataacc acagttacta cgtgggcatt attctcaaga 420 ccccaaataa caaatgagta cgtgaccagg gttccaatag actatcatga aattgtcagg 480 attgatcgat cgggagaatg ctcatccaaa gcaacgtatc ataaaaattt catgtttttt 540 gaagcttacg acaatgatga agcagaaaaa aaattgcccc tggttccatc actgttaaga 600 tcaactgtct ccaaggcgtt tcatacaact aactttacta agcgacatca aaccctggga 660 taccgaacgt ctacatcggt cgactgtgtt gtgaaatatc tacaggctag atctgtatac 720 ccgtatgatt actttggaat ggcgacaggt gatacagtag aaatttctcc tttttatacc 780 aaaaacacga ccggaccaag gcgtcacagt gtctacagag actatagatt tctcgaaatc 840 gcaaattatc aagtcaggga tttggaaacc ggacaaataa gaccccctaa aaaaagaaac 900 tttctaacag atgaacaatt cactataggc tgggatgcaa tggaagaaaa ggaatctgta 960 tgtactctca gtaaatggat tgaagtcccg gaagcagttc gtgtttcgta caaaaacagt 1020 taccactttt cacttaaaga tatgactatg acgttctcgt ccggaaaaca accttttaac 1080 atcagcaggc ttcatttggc tgaatgcgtt cctaccatag cctcggaggc catagatggc 1140 atctttgcca gaaagtatag ttcgactcat gtccgttctg gggacatcga atactatctc 1200 ggtagtggcg gatttctgat cgcatttcag aaactcatga gccatggctt ggctgaaatg 1260 tacctagaag aggcacaaag acaaaatcat ctcccgagag ggagagagcg tcgccaagcc 1320 gcaggtcgcc gcacggcgtc gctgcagtct ggacctcagg gtgatagaat tactacccac 1380 agttctgcaa catttgccat gttacaattt gcatacgaca aaatccaagc ccatgttaac 1440 gagcttatcg gaaatttgtt ggaagcgtgg tgtgagcttc agaaccgcca actgattgta 1500 tggcatgaga tgaagaaact aaacccgaac tcactgatga catctttgtt cggacaacct 1560 gtaagcgcca ggctattggg agacatcgta gcggtatcaa aatgtataga aattccaatc 1620 gaaaatatta ggatgcagga ttccatgcgc atgccagggg acccaaccat gtgctatacc 1680 agaccagtac ttattttcag gtattcgtcc tcccctgagt cacagttttc tgcgaactca 1740 acagaaaacc acaatcttga catattaggc caactcggag aacataatga aattttacaa 1800 gggcggaatt tgatagaacc atgcatgatc aatcacagac ggtactttct gttgggagaa 1860 aactaccttc tttacgaaga ctatacattt gttagacaag taaatgcttc cgagatcgaa 1920 gaagtgagca cattcatcaa cttgaacgcc actatactag aagatttgga ctttgtgccc 1980 gtcgaagtat acactcgcga ggaactcaga gatactggga ctttaaacta tgatgatgtg 2040 gtcagatatc aaaatattta taacaaaagg ttcagagaca ttgacactgt aatacgtgga 2100 gataggggag atgcaatctt tagagcaata gcagattttt ttggcaacac tcttggagaa 2160 gtaggaaagg cattgggaac tgtagtgatg acagccgcgg cagcagtaat ttctacagta 2220 tctggcatcg cctcatttct ttctaacccg ttcgccgcac tcggaattgg gatagcggtg 2280 gtggtgagca ttattttagg actgctggcg ttcaaatatg taatgaacct gaaatcaaac 2340 ccagttcagg ttctgttccc aggcgcagtt cccccggccg gaactcctcc acgaccctct 2400 agacgttact acaaggatga ggaggaggtt gaggaggata gtgatgagga cgacaggata 2460 cttgccacca gagttctgaa aggccttgag cttctacaca aggatgaaca gaaagctcga 2520 agacagaaag cgcggttttc tgcttttgct aaaaatatga gaaacctatt tcgcagaaaa 2580 ccccgaacca aggaagatga ctaccccctg ctcgaatacc cttcgtgggc agaagaaagc 2640 gaagacgaat aa 2652 <210> SEQ ID NO 14 <211> LENGTH: 1305 <212> TYPE: DNA <213> ORGANISM: Gallid herpesvirus 1 strain USDA <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1305) <223> OTHER INFORMATION: Laryngotracheitis virus (ILT) glycoprotein D <400> SEQUENCE: 14 atgcaccgtc ctcatctcag acggcactcg cgttactacg cgaaaggaga ggtgcttaac 60 aaacacatgg attgcggtgg aaaacggtgc tgctcaggcg cagctgtatt cactcttttc 120 tggacttgtg tcaggattat gcgggagcat atctgctttg tacgcaacgc tatggaccgc 180 catttatttt tgaggaatgc tttttggact atcgtactgc tttcttcctt cgctagccag 240 agcaccgccg ccgtcacgta cgactacatt ttaggccgtc gcgcgctcga cgcgctaacc 300 ataccggcgg ttggcccgta taacagatac ctcactaggg tatcaagagg ctgcgacgtt 360 gtcgagctca acccgatttc taacgtggac gacatgatat cggcggccaa agaaaaagag 420 aaggggggcc ctttcgaggc ctccgtcgtc tggttctacg tgattaaggg cgacgacggc 480 gaggacaagt actgtccaat ctatagaaaa gagtacaggg aatgtggcga cgtacaactg 540 ctatctgaat gcgccgttca atctgcacag atgtgggcag tggactatgt tcctagcacc 600 cttgtatcgc gaaatggcgc gggactgact atattctccc ccactgctgc gctctctggc 660 caatacttgc tgaccctgaa aatcgggaga tttgcgcaaa cagctctcgt aactctagaa 720 gttaacgatc gctgtttaaa gatcgggtcg cagcttaact ttttaccgtc gaaatgctgg 780 acaacagaac agtatcagac tggatttcaa ggcgaacacc tttatccgat cgcagacacc 840 aatacacgac acgcggacga cgtatatcgg ggatacgaag atattctgca gcgctggaat 900 aatttgctga ggaaaaagaa tcctagcgcg ccagaccctc gtccagatag cgtcccgcaa 960 gaaattcccg ctgtaaccaa gaaagcggaa gggcgcaccc cggacgcaga aagcagcgaa 1020 aagaaggccc ctccagaaga ctcggaggac gacatgcagg cagaggcttc tggagaaaat 1080 cctgccgccc tccccgaaga cgacgaagtc cccgaggaca ccgagcacga tgatccaaac 1140 tcggatcctg actattacaa tgacatgccc gccgtgatcc cggtggagga gactactaaa 1200 agttctaatg ccgtctccat gcccatattc gcggcgttcg tagcctgcgc ggtcgcgctc 1260 gtggggctac tggtttggag catcgtaaaa tgcgcgcgta gctaa 1305 <210> SEQ ID NO 15 <211> LENGTH: 1734 <212> TYPE: DNA <213> ORGANISM: Newcastle disease virus strain La Sota C5 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1734) <223> OTHER INFORMATION: HN gene <400> SEQUENCE: 15 atggaccgcg ccgttagcca agttgcgtta gagaatgatg aaagagaggc aaaaaataca 60 tggcgcttga tattccggat tgcaatctta ttcttaacag tagtgacctt ggctatatct 120 gtagcctccc ttttatatag catgggggct agcacaccta gcgatcttgt aggcataccg 180 actaggattt ccagggcaga agaaaagatt acatctacac ttggttccaa tcaagatgta 240 gtagatagga tatataagca agtggccctt gagtctccgt tggcattgtt aaatactgag 300 accacaatta tgaacgcaat aacatctctc tcttatcaga ttaatggagc tgcaaacaac 360 agtgggtggg gggcacctat ccatgaccca gattatatag gggggatagg caaagaactc 420 attgtagatg atgctagtga tgtcacatca ttctatccct ctgcatttca agaacatctg 480 aattttatcc cggcgcctac tacaggatca ggttgcactc gaataccctc atttgacatg 540 agtgctaccc attactgcta cacccataat gtaatattgt ctggatgcag agatcactca 600 cattcatatc agtatttagc acttggtgtg ctccggacat ctgcaacagg gagggtattc 660 ttttctactc tgcgttccat caacctggac gacacccaaa atcggaagtc ttgcagtgtg 720 agtgcaactc ccctgggttg tgatatgctg tgctcgaaag tcacggagac agaggaagaa 780 gattataact cagctgtccc tacgcggatg gtacatggga ggttagggtt cgacggccag 840 taccacgaaa aggacctaga tgtcacaaca ttattcgggg actgggtggc caactaccca 900 ggagtagggg gtggatcttt tattgacagc cgcgtatggt tctcagtcta cggagggtta 960 aaacccaatt cacccagtga cactgtacag gaagggaaat atgtgatata caagcgatac 1020 aatgacacat gcccagatga gcaagactac cagattcgaa tggccaagtc ttcgtataag 1080 cctggacggt ttggtgggaa acgcatacag caggctatct tatctatcaa ggtgtcaaca 1140 tccttaggcg aagacccggt actgactgta ccgcccaaca cagtcacact catgggggcc 1200 gaaggcagaa ttctcacagt agggacatct catttcttgt atcaacgagg gtcatcatac 1260 ttctctcccg cgttattata tcctatgaca gtcagcaaca aaacagccac tcttcatagt 1320 ccttatacat tcaatgcctt cactcggcca ggtagtatcc cttgccaggc ttcagcaaga 1380 tgccccaact cgtgtgttac tggagtctat acagatccat atcccctaat cttctataga 1440 aaccacacct tgcgaggggt attcgggaca atgcttgatg gtgtacaagc aagacttaac 1500 cctgcgtctg cagtattcga tagcacatcc cgcagtcgca ttactcgagt gagttcaagc 1560 agtaccaaag cagcatacac aacatcaact tgttttaaag tggtcaagac taataagacc 1620 tattgtctca gcattgctga aatatctaat actctcttcg gagaattcag aatcgtcccg 1680 ttactagttg agatcctcaa agatgacggg gttagagaag ccaggtctgg ctag 1734 <210> SEQ ID NO 16 <211> LENGTH: 1716 <212> TYPE: DNA <213> ORGANISM: Newcastle disease virus strain M08-3313/Mexico <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1716) <223> OTHER INFORMATION: Hemagglutinin neuramidase of Newcastle disease virus strain M08-3313/Mexico <400> SEQUENCE: 16 atggatcgtg tagttagcag agttgtacta gagaacgaag aaagagaagc aaagaacaca 60 tggcgcttgg ttttccggat cgcagtctta tctctaatag taatgacttt agctatctct 120 gtagccgccc tggtatacag catgggggct agcacaccga gcgaccttgc aggcatatca 180 acggtgatct ctaaggcaga ggatagggtt acatctttac tcagttcaaa tcaagatgtg 240 gtagataggg tatataaaca ggtggccctt gagtccccgc tggcgttgct aaatactgaa 300 tctataatta tgaatgcaat aacttctctc tcttatcaaa ttaatggggc tgcaaataat 360 agtgggtgtg gggcacctgt tcatgaccca gattatattg ggggggtagg caaagaactc 420 atagtagatg acacaagtga tgtcacatca ttctatcctt cagcatacca agaacacctg 480 aattttatcc cggcgcctac tacaggatca ggctgcactc ggataccctc attcgacatg 540 agcgctaccc actattgtta tactcacaat gtgatattat ctggttgcag agatcactcg 600

cactcacatc agtatttagc gctaggtgta cttcggacat ctgcaacagg gagggtattc 660 ttttctactc tgcgttccat caatttagat gacacccaaa atcggaagtc ttgcagtgtg 720 agtgcgactc ctttaggttg tgatatgctg tgctctaaag tcatagagac tgaggaggag 780 gattataagt caattacccc cacatcaatg gtgcatggaa ggttagggtt tgacggtcag 840 caccatgaga aggacttaga cgtcacagtc ttatttaagg attgggttgc aaattacccg 900 ggagtgggag gagggtctct tattgacgac cgtgtatggt tcccagttta tggagggcta 960 aaacccaatt cgcctagcga cactgcacaa gaagggagat atgtaatata caagcgctat 1020 aataacacat gccccgatgg acaagattac caagttcgga tggctaagtc ttcgtataag 1080 cctggacggt ttggtagaaa gcgcgtacag caagccatct tatctatcaa agtatcaaca 1140 tccttgggcg aggacccggt gctgactgta ccgccaaata cagttacact catgggggct 1200 gaaggcagag tcctcacagt agggacatct catttcttgt accaacgagg gtcttcatac 1260 ttctctcccg ccttattata ccctatgaca gtacacaaca aaacagctac tcttcatagt 1320 ccttatatat ttaatgcttt cactcggcca ggtagtgtcc cttgccaggc atcagcaagg 1380 tgccctaact catgtatcac tggagtctat actgatccgt atcctgtagt cttccatagg 1440 aatcacacct tgcgaggggt gttcgggaca atgcttgata atgaacaagc aaggttcaac 1500 cccgtatctg cagtatttga ttacacatct cgcagtcgca taacccgggt aagttcaagc 1560 agcaccaagg cagcatacac gacatcgaca tgttttaaag ttgtcaagac caataaagtt 1620 tattgcctta gcattgcaga aatatccaac accctatttg gggaattcag gattgtccct 1680 ttactggtcg agatcctcaa ggatgatagg gtttaa 1716 <210> SEQ ID NO 17 <211> LENGTH: 1350 <212> TYPE: DNA <213> ORGANISM: Chicken anemia virus isolate CAV-10 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1350) <223> OTHER INFORMATION: VP1 gene of Chicken anemia virus isolate CAV-10 <400> SEQUENCE: 17 atggcaagac gagctcgcag accgagaggc agattttacg ccttcagaag aggacggtgg 60 caccacctca agcgacttcg acgaagatat aaatttcgac atcggaggag acagcggtat 120 cgtagacgag cttttaggaa ggcctttcac aacccccgcc ccggtacgta tagtgtgagg 180 ctaccgaacc cccagtcaac gatgactatc cgctttcaag gcatcatttt tcttaccgaa 240 ggactcattc tacctaaaaa cagtacagcg gggggatatg cagaccacct ctacggggcg 300 agagtcgcca agatctcagt gaacctgaaa gagttcctgc tagcgtcaat gaacctgaca 360 tacgtgagca aaataggagg ccccatcgcc ggtgagttga ttgcggacgg gtctcaatca 420 caagccgcgc aaaactggcc taattgctgg ctgccgctag ataataacgt gccctccgct 480 acaccatcgg catggtggag atgggcttta atgatgatgc agcccacgga ctcttgccgg 540 ttctttaatc accctaaaca aatgaccctg caagacatgg gtcggatgtt tgggggctgg 600 catctgttcc gacacattga aacccgcttt cagctccttg ccactaagaa tgagggatcc 660 ttcagccccg tggcgagtct tctctcccag ggagagtacc tcacgcgtcg ggacgatgtt 720 aagtacagca gcgatcacca gaaccggtgg cgaaaaggcg agcaaccgat gacggggggt 780 attgcttatg cgacggggaa aatgagaccc gacgagcaac agtaccctgc tatgcccccc 840 gaccccccga tcatcacaac taccacaccg caaggcacgc aagtccgctg catgaatagc 900 acgcaagctt ggtggtcgtg ggacacatat atgagctttg caacactcac agcgctcggt 960 gcacaatggt ctttccctcc agggcaacgt tcagtttcta gacgctcgtt caaccaccac 1020 aaagcgagag gggccgggga ccccaaggga cagagatggc acacgcttgt tccgctcggc 1080 acagagacca tcaccgacag ctacatgagt gcgccggcat cagagctgga cactaacttc 1140 tttacgcttt acgtagcgca aggcacaaat aaatcgcagc agtacaagtt cggcacagct 1200 acatacgcgc taaaggaacc ggtaatgaag agcgatgcat gggcggtggt acgcgtccag 1260 tcagtctggc aactgggtaa caggcaaagg ccatacccat gggacgtcaa ctgggccaac 1320 agcaccatgt actgggggtc gcagccctga 1350 <210> SEQ ID NO 18 <211> LENGTH: 651 <212> TYPE: DNA <213> ORGANISM: Chicken anemia virus isolate CAV-10 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(651) <223> OTHER INFORMATION: VP2 gene of chicken anemia virus isolate CAV-10 <400> SEQUENCE: 18 atgcacggga acggcggaca accggccgct gggggcagtg aatcggcgct tagccgagag 60 gggcaacctg ggcccagcgg agccgcgcag gggcaagtaa tttcaaatga acgctctcca 120 agaagatact ccacccggac catcaacggt gttcaggcca ccaacaagtt cacggccgtt 180 ggaaacccct cactgcagag agatccggat tggtatcgct ggaattacaa tcactctatc 240 gctgtgtggc tgcgcgaatg ctcgcgctcc cacgctaaga tctgcaactg cggacaattc 300 agaaagcact ggtttcaaga atgtgccgga cttgaggacc gatcaaccca agcctccctc 360 gaagaagcga tcctgcgacc cctccgagta cagggtaagc gagctaaaag aaagcttgat 420 taccactact cccagccgac cccgaaccgc aagaaggtgt ataagactgt aagatggcaa 480 gacgagctcg cagaccgaga ggcagatttt acgccttcag aagaggacgg tggcaccacc 540 tcaagcgact tcgacgaaga tataaatttc gacatcggag gagacagcgg tatcgtagac 600 gagcttttag gaaggccttt cacaaccccc gccccggtac gtatagtgtg a 651 <210> SEQ ID NO 19 <211> LENGTH: 1482 <212> TYPE: DNA <213> ORGANISM: Fowl adenovirus 4 <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1482) <223> OTHER INFORMATION: Short fiber gene <400> SEQUENCE: 19 ggtcctatcc ctttttccta tcagggttac gtctactccc ccaacgggaa caatgctccg 60 ggcccctaaa agaagacatt ccgaaaacgg gaagcccgag accgaagcgg gaccttcccc 120 ggctccaatc aagcgcgcca aacgcatggt gagagcatcc cagcttgacc tggtttatcc 180 tttcgattac gtggccgacc ccgtcggagg gctcaacccg ccttttttgg gaggctcagg 240 acccctagtg gaccagggcg gacagcttac gctcaacgtc accgatccca tcatcatcaa 300 gaacagatcg gtggacttgg cccacgaccc cagtctcgat gtcaacgccc aaggtcaact 360 ggcggtggcc gttgaccccg aaggggccct ggacatcacc cccgatggac tggacgtcaa 420 ggtcgacccg tccggcggat tggattccac cgcgggtgga ctgggggtca gcgtggacga 480 caccttgctc gtggatcagg gagaactggg cgtacacctc aaccaacaag gacccatcac 540 tgccgatagc agtggtatcg acctcgagat caatcctaac atgttcacgg tcaacacctc 600 gaccggaagc ggagtgctgg aactcaacct aaaagcgcag ggaggcatcc aagccgacag 660 ttcgggagtg ggcgtttccg tggatgaaag cctacagatt gtcaacaaca ctctggaagt 720 gaaaccggat cccagcggac cgcttacggt ctccgccaat ggcctagggc tgaagtacga 780 cactaatacc ctagcggtga ccgcgggcgc tttaaccgtg gtcggagggg ggagcgtctc 840 cacacccaat cgctactttt gtctcgggaa gtcacagcct caacacctac aatgccacgc 900 cgtcaattcc agcgcgaacg ccttctcttg cgcctactac cttcaacagt ggaacataca 960 ggggctcctt gttacctccc tctacttgaa attggacagc gccaccatgg ggaatcgccc 1020 tggggacctc aactccgcca atgccaaatg gttcaccttt tgggtgtccg cctatctcca 1080 gcaatgcaac tccgggattc aagcgggaac ggtcagcccc tccaccgcca ccctcacgga 1140 ctttgaaccc atggccaata ggagcgtgac cagcccatgg acgtactcgg ccaatggata 1200 ctatgaacca tccatcgggg aattccaagt gttcagcccg gtggtaacag gtgcctggaa 1260 cccgggaaac atagggatcc gcgtcctccc cgtgccggtt tcggcctccg gagagcgata 1320 cacccttcta tgctatagtc tgcagtgcac gaacgcgagc atttttaatc caaacaacag 1380 cggaaccatg atcgtgggac ccgtgctcta cagctgtcca gcggcctccc tcccgtaagc 1440 gcgccctccc caccgcgtgt caaataaaga gtcatgaacg tt 1482 <210> SEQ ID NO 20 <211> LENGTH: 3510 <212> TYPE: DNA <213> ORGANISM: Infectious bronchitis virus <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(3510) <223> OTHER INFORMATION: Infectious bronchitis virus strain ArkDPI- derived commercial vaccine D spike glycoprotein gene <400> SEQUENCE: 20 atgttggtga agtcactgtt tctagtgacc attttgtttg cactatgtag tgctaattta 60 tatgacaacg aatcttttgt gtattactac cagagtgctt ttaggccagg acatggttgg 120 catttatatg gaggtgctta tgcagtagtt aatgtgtcta gtgaaaataa taatgcaggt 180 actgccccaa gttgcactgc tggtgctatt ggctacagta agaatctcag tgcggcctca 240 gtagccatga ctgcaccact aagtggtatg tcatggtctg ccaactcttt ttgtacagcc 300 cactgtaatt ttacttctta tatagtgttt gttacacatt gttataagag cggatctaat 360 agttgtcctt tgacaggtct tattccaagc ggttatattc gtattgctgc tatgaaacat 420 ggaagtgcta tgcctggtca cttattttat aatttaacag tttctgtgac taaatatcct 480 aagtttagat cgctacaatg tgttaataat tatacttctg tatatttaaa tggtgacctt 540 gttttcacat ctaactatac tgaagatgtt gtagctgcag gtgtccattt taaaagtggt 600 ggacctataa cttataaagt tatgagagag gttaaatcct tggcttattt tgtcaatggt 660 actgcacatg atgtcattct atgtgatgac acacctagag gtttgttagc atgccaatat 720 aatactggca atttttcaga tggcttctat ccttttacta atactagtat tgttaaggat 780 aagtttattg tttatcgtga aagtagtgtc aatactactt taacattaac taatttcacg 840 tttagtaatg aaagtggtgc ccctcctaat acaggtggtg ttgacagttt tattttatac 900 cagacacaaa cagctcagag tggttattat aattttaact tttcatttct gagtagtttt 960 gtttataggg aaagttatta tatgtatgga tcttaccatc cacgttgtag ttttagacct 1020 gaaaccctta ataatggttt gtggtttaat tccctttctg tttcattaac atacggtccc 1080 attcaaggtg gttgtaagca atctgtattt aatggtaaag caacttgttg ttatgcttat 1140 tcatacggag gacctcgtgg ttgtaaaggt gtctatagag gtgagctaac acagcatttt 1200 gaatgtggtt tgttagttta tgttactaag agcgatggct cccgtataca aactgcaaca 1260 caaccacctg tattaaccca aaatttttat aataacatca atttaggtaa gtgtgttgat 1320

tataatatat atggcagaat tggccaaggt cttattacta atgtaaccga cttagctgtt 1380 agttataatt atttatcaga cgcaggtttg gctattttag atacatctgg tgccatagac 1440 atcttcgttg tacaaggtga atatggtcct aactattata aggttaatcc atgtgaagat 1500 gtcaaccaac agtttgtagt ttctggtggt aaattagtag gtattctcac ttcacgtaat 1560 gaaacaggtt ctcagcttct tgagaaccag ttttatatta aaatcactaa tggaactcgt 1620 cgttctagac gttctgttac tgaaaatgtt acaaattgcc cttatgttag ttatggcaag 1680 ttttgtataa aacctgatgg ttcaatttct gtaatagtac caaaagaact ggatcagttt 1740 gtggcacctt tacttaatgt tactgaatat gtgctcatac ctaacagttt taatttaact 1800 gttacagatg agtacataca aacgcgtatg gataagatcc aaattaattg cctgcagtat 1860 gtttgtggca attctttggc ctgtagaaag ctgtttcaac aatatgggcc tgtttgtgac 1920 aacatattgt ctgtagtaaa tagtgttggt caaaaagaag atatggaact tttaaatttc 1980 tattcttcta ctaaaccagc tcgttttaat acaccagttt ttagtaatct tagtactggt 2040 gagtttaata tttctctttt gttaacaccc cctagtagtc ctaggaggcg ttcttttatt 2100 gaagatcttt tatttacaag tgttgaatct gtaggattac caacagatga cgcatacaaa 2160 atgtgcactg caggaccttt aggctttctt aaagaccttg catgtgctcg tgaatataat 2220 ggtttgcttg tgttgcctcc tattataaca gcagaaatgc aaactttgta tactagttct 2280 ttagtagctt ctatggcttt tggtggtatt actgcagctg gtgccatacc ttttgccaca 2340 caactgcagg ctagaattaa tcacttgggt attacccagt cacttttgtt gaagaatcaa 2400 gaaaaaattg ctgcttcctt taataaggcc attggtcata tgcaggaagg ttttaggagt 2460 acatctctag cattacaaca aattcaagat gttgttaata agcagagtgc tattcttact 2520 gagactatgg cagcacttaa taaaaatttt ggtgctattt cttctgtgat tcaagacatt 2580 taccagcaac ttgattccat acaagcagat gctcaagtgg atcggctcat aactggtaga 2640 ttgtcatcac tttctgtctt agcatctgct aagcagtcgg agtacattag agtgtcacaa 2700 cagcgtgagt tagctactca gaaaattaat gagtgtgtta aatcacagtc tattaggtat 2760 tccttttgtg gtaatggacg acatgtttta accataccac aaaatgcccc taatggtata 2820 gtgtttatac actttactta tacaccagag agctttatta atgttactgc aatagtgggt 2880 ttttgtgtaa gtcctgctaa tgctagtcag tatgcaatag tgcccgctaa tggtaggggt 2940 atttttatac aagttaatgg tagttactac atcactgcac gagatatgta tatgccaaga 3000 gatattactg caggagatat agttacgctt acttcttgtc aagcaaatta tgtaagtgta 3060 aataagaccg tcattactac atttgtagac aatgatgatt ttgattttgc tgatgaattg 3120 tcaaaatggt ggaatgatac taagcatgag ctaccagact ttgacaaatt caattacaca 3180 gtacctatac ttgacattga tagtgaaatt gatcgtattc aaggcgttat acagggtctt 3240 aacgactctc taatagacct tgaaacacta tcaatactca aaacttatat taagtggcct 3300 tggtatgtgt ggttagccat agcttttgcc actattatct tcatcttaat actaggatgg 3360 ttgtttttca tgactggttg ttgtggttgt tgttgtggat gctttggcat tattccttta 3420 atgagtaagt gtggtaagaa atcttcttat tacacgactt ttgataatga tgtggtaact 3480 gaacaataca gacctaaaaa gtctgtttaa 3510 <210> SEQ ID NO 21 <211> LENGTH: 1617 <212> TYPE: DNA <213> ORGANISM: Avian metapneumovirus <220> FEATURE: <221> NAME/KEY: gene <222> LOCATION: (1)..(1617) <223> OTHER INFORMATION: Gene F of Avian metapneumovirus isolate IT/Ty/A/259-01/03 <400> SEQUENCE: 21 atggatgtaa gaatctgtct cctattgttc cttatatcta atcctagtag ctgcatacaa 60 gaaacataca atgaagaatc ctgcagtact gtaactagag gttataagag tgtgttaagg 120 acagggtggt atacgaatgt atttaacctc gaaataggga atgttgagaa catcacttgc 180 aatgatggac ccagcctaat tggcactgag ttagtactca caaagaatgc tttgagggag 240 ctcaaaacag tgtcagctga tcaagtggct aaggaaagca gactatcctc acccaggaga 300 cgtagatttg tactgggtgc aattgcactt ggtgttgcga cagctgctgc cgtaacagct 360 ggtgtagcac ttgcaaagac aattagatta gagggagagg tgaaggcaat taagaatgcc 420 ctccggaaca caaatgaggc agtatccaca ttagggaatg gtgtgagggt actagcaact 480 gcagtcaatg acctcaaaga atttataagt aaaaaattga ctcctgctat taaccagaac 540 aaatgcaata tagcagatat aaagatggca attagttttg gccaaaataa cagaaggttc 600 ctgaatgtgg tgaggcaatt ctctgatagt gcaggtatca catcagctgt gtctcttgat 660 ttaatgacag atgatgaact tgttagagca attaacagaa tgccaacttc atcaggacag 720 attagtttga tgttgaacaa tcgtgccatg gttagaagga aggggtttgg tatattgatt 780 ggtgtttatg atggaacggt cgtttatatg gtacaactgc ccatattcgg cgtgattgag 840 acaccttgtt ggagggtggt ggcagcacca ctctgtagga aaaagaaagg caattatgct 900 tgtatactga gagaagatca agggtggtac tgtacaaatg ctggctctac agcttattat 960 cctaatgaag atgattgtga ggtaagggat gattatgtat tttgtgacac agcagctggc 1020 attaatgtgg ccctagaagt tgaacagtgc aactataaca tatcgacttc taaataccca 1080 tgcaaagtca gcacaggtag acaccctgtc agtatggtag ccttaacccc cctagggggt 1140 ctagtgtctt gttatgagag tgtaagttgc tccataggta gcaataaagt agggataata 1200 aaacagctag gcaaagggtg cacccacatt cccaacaacg aagctgacac gataaccatt 1260 gataacactg tgtaccaatt gagcaaggtt gtaggcgaac agaggaccat aaaaggagct 1320 ccagttgtga acaattttaa cccaatatta ttccctgagg atcagttcaa tgttgcactt 1380 gaccaagtat ttgagagtat agatagatct caggacttaa tagataagtc taacgacttg 1440 ctaggtgcag atgccaagag caaggctgga attgctatag caatagtagt gctagtcatt 1500 ctaggaatct tctttttact tgcagtgata tattactgtt ccagagtccg gaagaccaaa 1560 ccaaagcatg attacccggc cacgacaggt catagcagca tggcttatgt cagttaa 1617 <210> SEQ ID NO 22 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer pCI-Neo <400> SEQUENCE: 22 ggctcatgtc caatatgacc gccat 25 <210> SEQ ID NO 23 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer EGFP <400> SEQUENCE: 23 aaaaaagaat tcaccatggt gagcaagggc gag 33 <210> SEQ ID NO 24 <211> LENGTH: 38 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer EGFP <400> SEQUENCE: 24 aaaaaagcgg ccgcttactt gtacagctcg tccatgcc 38 <210> SEQ ID NO 25 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer EGFP-I <400> SEQUENCE: 25 gagaagcgcg atcacatggt 20 <210> SEQ ID NO 26 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer beta-actin <400> SEQUENCE: 26 aaaaaaacta gttggtcgag gtgagcccca cgtt 34 <210> SEQ ID NO 27 <211> LENGTH: 32 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer beta actin <400> SEQUENCE: 27 aaaaaagaat tcgcccgccg cgcgcttcgc tt 32 <210> SEQ ID NO 28 <211> LENGTH: 44 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer L2R <400> SEQUENCE: 28 aaaaaaacta gtcatagtaa atatgggtaa cttcttaata gcca 44 <210> SEQ ID NO 29 <211> LENGTH: 45 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer L2R <400> SEQUENCE: 29 aaaaaagaat tcatggtttg ttattggcaa atttaaatta atgtt 45 <210> SEQ ID NO 30 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer Luc <400> SEQUENCE: 30 aaaaaaccta ggttggcaat ccggtactgt tggt 34 <210> SEQ ID NO 31

<211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer Luc <400> SEQUENCE: 31 aaaaaattcg aagccgcccc gactctag 28 <210> SEQ ID NO 32 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer SV40 <400> SEQUENCE: 32 ggcctctgag ctattccaga 20 <210> SEQ ID NO 33 <211> LENGTH: 42 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer WPRE <400> SEQUENCE: 33 aaaaaagcgg ccgctcaacc tctggattac aaaatttgtg aa 42 <210> SEQ ID NO 34 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer WPRE <400> SEQUENCE: 34 aaaaaagcgg ccgcgcccaa agggagatcc gac 33 <210> SEQ ID NO 35 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer VF1 <400> SEQUENCE: 35 aaaaaaagat cttacatgaa tgacgctgct g 31 <210> SEQ ID NO 36 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer VF1 <400> SEQUENCE: 36 aaaaaaacta gtaaatacac cgagaaatac cacg 34 <210> SEQ ID NO 37 <211> LENGTH: 38 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer VF2 <400> SEQUENCE: 37 aaaaaacaat tgaaataaag actcagaacg cattttcc 38 <210> SEQ ID NO 38 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer VF2 <400> SEQUENCE: 38 aaaaaaggta cccccccgca gagaattaaa a 31 <210> SEQ ID NO 39 <211> LENGTH: 18 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer E1 <400> SEQUENCE: 39 acatgaatga cgctgctg 18 <210> SEQ ID NO 40 <211> LENGTH: 19 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer E1 <400> SEQUENCE: 40 cccccgcaga gaattaaaa 19 <210> SEQ ID NO 41 <211> LENGTH: 496 <212> TYPE: DNA <213> ORGANISM: Avian encephalomyelitis virus <220> FEATURE: <221> NAME/KEY: misc_feature <222> LOCATION: (1)..(496) <223> OTHER INFORMATION: Internal Ribosomal Entry Site (IRES) from a Canadian isolate of Avian encephalomyelitis virus <400> SEQUENCE: 41 tttgaaagag gcctccgggg tgtccggagg ttctctttcg acccaaccca tactgggggg 60 tgtgtggggc cgtatttgga gtgcacggta tatatgcatt cccgcatggc aagggcgtgc 120 caccttgccc cttgacgcat ggtatgtgtc attatttgcc ttggttaaac cccataaaac 180 gaggcgccac gtgccgaaaa tccctttgcg tttcacagaa ccatcctaac catgggtgta 240 gtatgggaat cgtgtatggg ggtgattagg atctctcgta gagggatagg tgtatcattc 300 aaatccaggg agtactctgg ctctgacatt gggacatttg atgtaaccgg acctggttca 360 gtatccgggt tgtcctgtat tgttacggtg tatccgtctt gacacactga aagggtattc 420 ttgggtaatc ccttcctact gcctgatagg gcggcgtgcc cggccacgag agattaaggg 480 tagcaattta aacatg 496 <210> SEQ ID NO 42 <211> LENGTH: 18 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer ORF1-2Ver <400> SEQUENCE: 42 gcacagtccc aatggctt 18 <210> SEQ ID NO 43 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer ORF1-2Ver <400> SEQUENCE: 43 aaattctggt ccgttaccga 20 <210> SEQ ID NO 44 <211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer TR2-17-11Ver <400> SEQUENCE: 44 gcctactcct cagcctatca c 21 <210> SEQ ID NO 45 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer TR2-17-11Ver <400> SEQUENCE: 45 cagtcctcta acgagcacgc 20 <210> SEQ ID NO 46 <211> LENGTH: 20 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer CAT <400> SEQUENCE: 46 gtgtaggctg gagctgcttc 20 <210> SEQ ID NO 47 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer ORF1CATSwaI <400> SEQUENCE: 47 ggacgtgctt taggtaattt cctccg 26 <210> SEQ ID NO 48 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 48 ttctttttcc cgtttaggtg aggag 25 <210> SEQ ID NO 49 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 49 atttaaatgt gtaggctgga gctgcttc 28 <210> SEQ ID NO 50 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence

<220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer ORF2CATSwaI <400> SEQUENCE: 50 gcgttctgag tctttattgg taa 23 <210> SEQ ID NO 51 <211> LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 51 actcgaaaca cgcgtcacac gcgc 24 <210> SEQ ID NO 52 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 52 atttaaatca tatgaatatc ctccttagtt c 31 <210> SEQ ID NO 53 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer TR2-17-11CAT <400> SEQUENCE: 53 ccccctggcg gccgttggcc gccactg 27 <210> SEQ ID NO 54 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 54 cacccgcgcc ggacttagtc tct 23 <210> SEQ ID NO 55 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 55 atttaaatgt gtaggctgga gctgcttc 28 <210> SEQ ID NO 56 <211> LENGTH: 27 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer TR2-17-11CAT <400> SEQUENCE: 56 agtagagatt ataggaagcg gggatta 27 <210> SEQ ID NO 57 <211> LENGTH: 23 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 57 tagtggtttt tatttaaacg aga 23 <210> SEQ ID NO 58 <211> LENGTH: 31 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 58 atttaaatca tatgaatatc ctccttagtt c 31 <210> SEQ ID NO 59 <211> LENGTH: 25 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer ORF17CATSwaI <400> SEQUENCE: 59 tccccctggc ggccgagggc cgcca 25 <210> SEQ ID NO 60 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 60 ctgcacccgc gccggactta gtctct 26 <210> SEQ ID NO 61 <211> LENGTH: 28 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer <400> SEQUENCE: 61 atttaaatgt gtaggctgga gctgcttc 28 <210> SEQ ID NO 62 <211> LENGTH: 26 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer ORF17CATSwaI <400> SEQUENCE: 62 gatacgaaga aggataggag cagaat 26 <210> SEQ ID NO 63 <211> LENGTH: 22 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 63 cgaggatacg gtagttgact cc 22 <210> SEQ ID NO 64 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer <400> SEQUENCE: 64 atatttaaat catatgaata tcctccttag ttc 33 <210> SEQ ID NO 65 <211> LENGTH: 34 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer EGFPcaSwaI <400> SEQUENCE: 65 agctgcattt aaatgtatta ccgccatgca ttag 34 <210> SEQ ID NO 66 <211> LENGTH: 33 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer EGFPcaSwaI <400> SEQUENCE: 66 agctgcattt aaatccacaa ctagaatgca gtg 33 <210> SEQ ID NO 67 <211> LENGTH: 36 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Forward primer mCherry <400> SEQUENCE: 67 agctgcattt aaatatggtg agcaagggcg aggagg 36 <210> SEQ ID NO 68 <211> LENGTH: 39 <212> TYPE: DNA <213> ORGANISM: Artificial Sequence <220> FEATURE: <223> OTHER INFORMATION: Synthetic - Reverse primer mCherry <400> SEQUENCE: 68 agctgcattt aaatctactt gtacagctcg tccatgccg 39

Claims

1. A fowl adenovirus 9 (FAdV-9) recombinant viral vector.

2. The FAdV-9 vector of claim 1, wherein the vector comprises one or more deletions at the left end and/or right end of the FAdV-9 genome.

3. The FAdV-9 vector of claim 1 or 2, wherein the vector comprises a deletion of one or more of ORF0, ORF1 and ORF2.

4. The FAdV-9 viral vector of any one of claims 1 to 3, wherein the vector comprises a deletion of one or more of ORF19, TR2, ORF17 and ORF11.

5. The FAdV-9 viral vector of any one of claims 1 to 4, wherein the vector comprises a deletion of ORF0, ORF1, ORF2, TR2, ORF17 and ORF11.

6. The FAdV-9 viral vector of any one or claims 1 to 5, wherein the vector has an insert capacity of greater than 4000 bp, greater than 5000 bp, greater than 6000 bp and optionally greater than 7000 bp.

7. The FAdV-9 viral vector of claim 1 or 2, wherein the vector comprises the nucleotide sequence shown in SEQ ID NO: 1 with one or more deletions, optionally one or more deletions shown in FIG. 10a.

8. The FAdV-9 viral vector of any one of claims 1 to 7, wherein the vector comprises at least one of the following: (a) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 575 to 2753 have been deleted, (b) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 847 to 2753 have been deleted, (c) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 42,398 have been deleted, (d) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 34,220 to 36,443 have been deleted, (e) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 40,561 have been deleted or wherein nucleotides 41,461 to 42,398 have been deleted, and (f) a nucleotide sequence with at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity to the nucleotide sequences set out in (a), (b), (c), (d) or (e).

9. The FAdV-9 viral vector of claim 8, wherein the vector comprises: (a) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 575 to 2753 have been deleted, and (b) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 42,398 have been deleted.

10. The FAdV-9 viral vector of claim 8, wherein the vector comprises: (a) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 847 to 2753 have been deleted, and (b) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 42,398 have been deleted.

11. The FAdV-9 viral vector of claim 8, wherein the vector comprises: (a) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 847 to 2753 have been deleted, and (b) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 34,220 to 36,443 have been deleted.

12. The FAdV-9 viral vector of claim 8, wherein the vector comprises: (a) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 847 to 2753 have been deleted, (b) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 34,220 to 36,443 have been deleted, and (c) a nucleotide sequence with sequence identity to the sequence shown in SEQ ID NO: 1, wherein nucleotides 38,807 to 40,561 have been deleted or wherein nucleotides 41,461 to 42,398 have been deleted.

13. The FAdV-9 viral vector of any one of claims 1 to 12, comprising one or more exogenous nucleotide sequences coding for one or more polypeptides of interest.

14. The FAdV-9 viral vector of claim 13, wherein the vector is a dual vector comprising two exogenous nucleotide sequences coding for two polypeptides of interest.

15. The FAdV-9 viral vector of any one or claims 1 to 14, comprising an exogenous nucleotide sequence coding for at least one antigenic site of a disease of concern.

16. The FAdV-9 viral vector of claim 15, wherein the exogenous nucleotide sequence is selected from antigenic sequences against influenza, infectious laryngotracheitis, infectious bronchitis, infectious bursal disease (Gumboro), hepatitis, viral rhinotracheitis, infectious coryza, Mycoplasma hyopneumonieae, pasteurellosis, Porcine Respiratory and Reproductive Syndrome (PRRS), circovirus, bordetellosis, parainfluenza and any other antigen which size allows its insertion into the corresponding viral vector.

17. The FAdV-9 viral vector of claim 16, wherein the exogenous nucleotide sequence is selected from antigenic sequences against Avian influenza, Laryngotracheitis (LT), Newcastle disease (NDV), infectious anemia, Inclusion bodies hepatitis, Infectious Bronchitis (IB), Metapneumovirus (MPV) and Gumboro.

18. The FAdV-9 viral vector of claim 16, wherein the exogenous nucleotide sequence comprises a sequence corresponding to at least one gene listed in Table 1 or a homolog thereof.

19. The viral vector of any one of claims 13 to 18, wherein the exogenous nucleotide sequence is operably linked to a control sequence, optionally a promoter sequence.

20. A host cell comprising the viral vector of any one of claims 1 to 19.

21. A method for producing the viral vector of any of claims 13 to 19, comprising the steps of: a) optionally amplifying the exogenous nucleotide sequence; b) inserting the exogenous nucleotide sequence in the viral vector; and, c) introducing the infectious clone thus produced into a suitable cell line.

22. The method of claim 21, wherein the exogenous nucleotide sequence is selected from antigenic sites sequences against influenza, infectious laryngotracheitis, infectious bronchitis, bursa of Fabricius' infection (Gumboro), hepatitis, viral rhinotracheitis, infectious coryza, Mycoplasma hyopneumonieae, pasteurellosis, Porcine Respiratory and Reproductive Syndrome (PRRS), circovirus, bordetellosis, parainfluenza, and any other antigen which size allows its insertion into the corresponding viral vector.

23. The method of claim 21, wherein the exogenous nucleotide sequence is selected from antigenic sites sequences against Avian influenza, Laryngotracheitis (LT), Newcastle disease (NDV), infectious anemia, Inclusion bodies, Infectious Bronchitis (IB), Metapneumovirus (MPV) and Gumboro.

24. The method of claim 21, wherein the exogenous nucleotide sequence comprises a sequence corresponding to at least one gene listed in Table 1 or a homolog thereof.

25. An immunogenic composition comprising at least the FAdV-9 viral vector of any of claims 1 to 19 with an exogenous nucleotide sequence coding for at least one antigenic site of a disease of concern inserted therein.

26. The immunogenic composition of claim 25, further comprising a pharmaceutically acceptable carrier.

27. The immunogenic composition of claim 24 or 25, further comprising an adjuvant.

28. A method of generating an immunogenic response in a subject, the method comprising administering to the subject the immunogenic composition of any one of claims 25 to 27.